Sidt2 yog ib qho tseem ceeb ntawm cov protein nyob rau hauv Autophagy-lysosomal degradation txoj kev thiab yog qhov tseem ceeb rau kev saib xyuas ntawm lub raum cov qauv thiab kev ua haujlwm lim
Nov 07, 2023
Kev tswj hwm thiab homeostasis ntawm autophagy yog qhov tseem ceeb rau kev tswj hwm lub cev morphology thiab kev ua haujlwm. Raws li lysosomal membrane protein, cov nyhuv ntawmSidt 2ntawmlub raum qauvthiablub raum autophagytseem tsis paub. Hauv txoj kev tshawb no, peb pom tias lub raum ntawmSidt2-/-Cov nas pom kev hloov pauv hauv qab daus daim nyias nyias, cov txheej txheem ko taw fusion, thiab mitochondrial o, tawm tswv yim tiastus qauv ntawm lub raum puas lawm. Cov zis muaj protein ntau nyob rau ntawm 24 teev qhia tias covlub raum ua haujlwmkuj raug puas tsuaj. Nyob rau tib lub sijhawm, qhov tsis muajSidt 2Ua rau txo qis ntawm cov kua qaub lysosomes, txo qis hauv cov kua qaub hydrolase kev ua haujlwm thiab kev qhia hauv lysosome, thiab nce pH hauv lysosome, qhia tias lysosomal ua haujlwm tsis zoo tom qab.Sidt 2kev tshem tawm. Kev sib sau ntawm autophagolysosomes, nce LC3-II thiab P62 protein ntau ntau, thiab txo qis P62 mRNA qib qhia tau hais tias tsis muajSidt 2gene ua rau txawv txav autophagy txoj kevflow ua. Kev sim tshuaj chloroquine, immunofluorescence autophagosome, lysosome fusion assay, thiab Ad-mcherry-GFP-LC3B ntxiv qhia tias, tom qabSidt 2deletion, zus tau tej cov autophagosomes tsis tau nce, tab sis fusion ntawm autophagosomes thiab lysosomes thiab degradation ntawm autophagolysosomes yog impaired. Thaum incubatingSidt2-/-cov hlwb nrog autophagy activator rapamycin, peb pom tias nws tuaj yeem ua rau autophagy, uas tshwm sim raws li kev nce hauv autophagosomes, tab sis nws tsis tuaj yeem txhim kho autophagolysosome degradation. Meanwhile, nws ntxiv illustrated tias lubSidt 2gene plays lub luag haujlwm tseem ceeb hauv kev ua tiav ntawm cov txheej txheem autophagolysosome. Hauv kev xaus, qhov tsis muajSidt 2Cov noob ua rau muaj kev cuam tshuam lysosome kev ua haujlwm thiab txo qis ntawm cov kua qaub lysosomes, ua rau kev tsim thiab degradation ntawm autophagolysosomes, uas nws thiaj li tshwm sim raws lilub raum txawv txav thiab kev ua haujlwm. Sidt 2Nws yog ib qho tseem ceeb hauv kev tswj hwm kev ua haujlwm ntawm lysosomes thiab kev ruaj ntseg ntawm lub raum.
Taw qhia
Qhov kev pom ib txwm muaj yog tias lysosomes yog lub hlwb cov khib nyiab pov tseg kom tshem tawm cov khib nyiab uas tsim los ntawm cov hlwb [1–3]. Txawm li cas los xij, ntau cov kev tshawb fawb tau pom tias lub luag haujlwm ntawm lysosomes yog qhov nyuaj dua, thiab tuaj yeem cuam tshuam txog kev hloov pauv ntawm tes, qog nqaij hlav, kev loj hlob, thiab lwm yam uas cuam tshuam rau lub neej ua haujlwm ntawm lub cev [4-10]. Lysosomes feem ntau enriched hauv cov ntaub so ntswg xws li lub siab thiab lub raum [11]. Yog li ntawd, lysosomal dysfunction kuj tseem cuam tshuam nrog cov kab mob ntawm daim siab thiab lub raum, xws li Gaucher's disease [12], mucopolysaccharidosis [13], Niemann-Pick kab mob [14], qeeb glomerulosclerosis [15], idiopathic membranous nephropathy [16] , thiab lwm yam. Lysosomal membrane proteins (LMPs) yog cov khoom siv membrane uas nws txoj haujlwm tsis yog tsuas yog ua kom muaj kev ncaj ncees ntawm lysosome tab sis kuj tseem koom nrog ntau yam xws li intracellular signal transduction thiab kev tswj hwm uas yog qhov tseem ceeb rau kev tswj hwm kev ua haujlwm ntawm lysosome. thiab kev ua ub no ntawm tes [17–19].
Txog niaj hnub no, ntau dua 100 LMPs tau pom, tab sis cov haujlwm ntawm lawv feem ntau tseem tsis tau paub [20]. Transmembrane 7 superfamily tus tswv cuab 1 (TM7SF1) yog qhov tseem ceeb rau kev saib xyuas ntawm lub raum podocyte muaj nuj nqi [21] thiab ua lub luag haujlwm tseem ceeb hauv cov txheej txheem ntawm lub raum kev loj hlob [22]. Chloride Voltage-Gated Channel 5 (ClC{10}}) yog chloride ion (Cl(−)) channel qhia nyob rau hauv lub raum tubules, uas yog ib qho tseem ceeb rau lub raum tubular muaj nuj nqi [23]; Thaum nws hloov pauv, nws yuav ua rau Dent's kab mob [24]. Overexpression ntawm Chloride Voltage-Gated Channel 7 (ClC-7) tiv thaiv apoptosis ntawm lub raum tubular epithelial hlwb tshwm sim los ntawm impaired redox xeev [25]. Cov kev tshawb fawb no qhia tau hais tias, raws li cov khoom ntawm lysosome-enriched raum cov ntaub so ntswg, LMPs yog qhov tseem ceeb los tswj kev ua haujlwm ib txwm muaj, tab sis cov txheej txheem tshwj xeeb ntawm lawv cov kab mob cuam tshuam tseem tsis meej.
SID1 transmembrane tsev neeg, tus tswv cuab 2 (Sidt2) yog ib qho LMP uas tau tshawb pom tshiab uas tau nthuav tawm hauv daim siab thiab raum cov ntaub so ntswg [26]. Cov kev tshawb fawb yav dhau los tau pom tias Sidt2 tshem tawm tuaj yeem ua rau muaj kab mob siab, uas tshwm sim raws li daim siab steatosis thiab daim siab lipid metabolism tsis zoo [11, 18]. Hauv kev tshawb fawb tsis ntev los no, peb pom tias ob lub raum ntawm Sidt2-/- nas kuj tau ntsib kev puas tsuaj, tab sis cov txheej txheem tshwj xeeb tsis meej. Lysosomes yog ib qho tseem ceeb thawj coj orga nelles rau autophagy [10]. Puas yog kev cai autophagy koom nrog? Hauv txoj kev tshawb no, peb tau tshuaj xyuas qhov kev sib raug zoo ntawm kev ua haujlwm ntawm lysosomal (autophagy) thiab kab mob thiab tshawb nrhiav cov txheej txheem hauv qab uas muaj Sidt2 uas ua rau lub raum puas, uas yuav muaj txiaj ntsig zoo rau kev kawm txog kev sib raug zoo ntawm LMP thiab kab mob.
TSEEM CEEB
Sidt2-/- qauv qhia tiasraum puasyog txuam nrog autophagolysosome tsub zuj zuj
Tus txheej txheem ntawm kev tsim Sidt2−/− nas yog qhia hauv daim duab 1A. Qhov tau txais homozygous nas tail DNA tau ua ntu zus thiab cov txiaj ntsig sib luag yog unimodal (Fig. 1B), thiab 199 bp fragment poob tshwm sim hauv qhov thib ob exon. Los ntawm PCR genotype txheeb xyuas, peb pom tias Sidt2+/+ (Wild Type, WT) nas muaj 685 bp DNA ntu, hos Sidt2−/− nas muaj 486 bp (Fig. 1C). Raws li pom hauv daim duab 1D, Sidt2 protein tsis tuaj yeem kuaj pom hauv Sidt2−/− nas dua li hauv WT, uas qhia tias tus qauv tau ua tiav. Los ntawm kev kuaj zis, nws tau pom tias 24 teev cov zis protein tau nce ntau hauv Sidt2−/− nas piv nrog WT (Fig. 1E), qhia tias lub raum pom kev tsis zoo tom qab Sidt2 raug tshem tawm. Kev sib kis ntawm electron microscopy kev soj ntsuam (Daim duab 1F) pom tias, piv nrog WT nas (a–f), Sidt2−/− nas ob lub raum nthuav tawm diffuse fusion ntawm ko taw txheej txheem, thickening ntawm glomerular hauv qab daim nyias nyias (g, h), raum tubular epithelial cell edema, microvilli puas (i, j), mitochondrial edema, vacuole-zoo li hloov, thiab ploj ntawm spines (k, l). Interestingly, Sidt2−/− nas kuj tau nthuav tawm ntau qhov autophagolysosome accumululations (pom liab, m, n), thiab tus naj npawb tau ntau dua li qhov kev tswj hwm (Daim duab 1G). Hauv qib cellular, peb pom tias Sidt2 tseem yog qhov tseem ceeb rau kev ciaj sia ntawm lub raum hlwb. Kev tshem tawm ntawm Sidt2 noob ua rau txo qis kev loj hlob thiab nce apoptosis ntawm MPC5 thiab SV40 MES 13 hlwb (Cov Lus Qhia Ntxiv 1).
Sidt2 noob tshem tawm ua rau muaj kev hloov pauv ntawm tus lej thiab kev ua haujlwm ntawm acidic lysosomes hauv nas raum hlwb
Crispr-Cas9 thev naus laus zis tau siv los rhuav tshem Sidt2 noob hauv MPC5 thiab SV40 MES 13 hlwb kom tau txais cov qauv ntawm tes. Kev kuaj xyuas mRNA thiab qib protein (Fig. 2A–C) tau ua los ntawm qRT PCR thiab western blotting raws li qhov qhia tau tias cov qauv tau ua tiav. Lub lysosome-mediated degradation system yog ib kauj ruam tseem ceeb hauv autophagy degradation. Raws li ib qho tseem ceeb ntawm LMP, puas yuav lub khob tawm ntawm Sidt2 cuam tshuam rau cov lus qhia ntawm lysosome-txog cov proteins? Peb ntsuas qhov loj LMP lysosomal-associated membrane protein 1 (LAMP1). Cov txiaj ntsig tau pom tias qhov kev qhia ntawm LAMP1 txo qis tom qab Sidt2 tshem tawm hauv MPC5 thiab SV40 MES 13 hlwb (Fig. 2D, E). Peb txuas ntxiv siv LysoTracker los sau cov kua qaub lysosomes thiab pom tias cov kua qaub lysosomes txo qis tom qab Sidt2 tshem tawm hauv ob hom hlwb (Fig. 2F, G). Tom qab ntawd, peb ntsuas lysosomal cathepsin B (CTSB) thiab pom tias nyob rau hauv nas raum cov ntaub so ntswg (Fig. 2H, I), MPC5 hlwb (Fig. 2J, K) thiab SV40 MES 13 hlwb (Fig. 2L, M), cov kab lus. ntawm CTSB hauv Sidt2-/- qauv tau txo qis, qhia tias kev ua haujlwm ntawm cov enzyme proteolytic hauv lysosomes tau txo qis thaum Sidt2 noob raug tshem tawm. Ib yam li ntawd, kev qhia ntawm tus thawj CTSB hauv ob hom hlwb ntawm Sidt2-/- pab pawg tau txo qis, qhia tias CTSB kuj tau cuam tshuam tom qab Sidt2 tshem tawm. Peb txuas ntxiv siv LysoSensor txhawm rau txheeb xyuas qhov acidic ib puag ncig ntawm lysosomes. Qhov qis dua qhov fluorescence siv piv, qhov pH ntau dua hauv lysosome. Cov txiaj ntsig tau pom tias tom qab Sidt2 tshem tawm, pH tus nqi nce ntxiv (Daim duab 2N, O) thiab acidification yog txawv txav. Txhawm rau tshawb xyuas ntxiv seb qhov lysosomal abnormalities saum toj no yog vim li cas cov lej ntawm lysosomes los yog txawv txav lysosomal functions, peb tshawb xyuas cov hlwb los ntawm electron microscopy (Fig. 2P). Peb pom tias tsis muaj qhov pom tseeb hloov pauv ntawm tus lej ntawm thawj lysosomes thiab tag nrho cov lysosomes (xws li thawj thiab theem nrab lysosomes) tom qab Sidt2 tshem tawm (Daim duab 2Q).
Lub raum autophagy txoj kev yog txawv txav tom qab Sidt2 tshem tawm
Raws li tau hais los saum no, ntau qhov autophagolysosome accumulations tau pom nyob rau hauv lub raum hlwb ntawm Sidt2-/- nas, qhia tias txoj kev autophagy yog txawv txav. Siv Western blot tsom xam, nws tau pom tias muaj qhov nce qib protein ntau ntawm LC3-phosphatidylethanolamine conjugate (LC3-II) thiab Sequestosome 1(P62) hauv lub raum ntawm Sidt2−/− nas, thiab qhov nce hauv autophagy-related protein qhia ntawm Autophagy ntsig txog 5(Atg5), Autophagy-txog 7(Atg7), thiab Autophagy-txog 12(Atg12) (Fig. 3A, B). Txawm li cas los xij, qib P62 mRNA tau txo qis (Daim duab 3C), qhia tias txoj kev autophagy yog qhov txawv txav tom qab tshem tawm Sidt2 hauv vivo. Tom qab ntawd peb pom tias cov kab lus ntawm autophagy proteins LC3-II, P62, Atg5, Atg7, thiab Atg12 kuj tau nce ntxiv (Daim duab 3D–G) hauv ob hom kab mob raum tom qab Sidt2 tshem tawm, uas zoo ib yam. nrog vivo. Qhov nce hauv LC3-II qhia tau tias muaj kev nce hauv autophagosomes hauv lub raum hlwb tom qab Sidt2 tshem tawm, uas tej zaum yuav tshwm sim los ntawm kev ua kom autophagy lossis blocked degradation ntawm autophagosomes. P62 immunofluorescence (Fig. 3H) qhia meej tias qhov nce ntawm P62 cov ntsiab lus hauv Sidt2−/− cells (Fig. 3I). Contrary to the protein level, qRT-PCR qhia tau hais tias P62 txo qis heev ntawm mRNA theem (Fig. 3J), qhia tias qhov nce ntawm P62 muaj feem xyuam rau tsis txaus degradation. Qhov nce hauv Atg5, Atg7, thiab Atg12 tau qhia tias autophagy pib thiab tsim ib txwm, tab sis kev sib sau ntawm P62 kuj tau qhia txog cov txheej txheem autophagy, thiab qhov kev tsis sib haum xeeb no xav kom peb los tshawb txog qhov tseeb ntawm autophagy flux.
Hauv vitro, chloroquine daim ntawv thov qhia txo qis autophagy flux tom qab Sidt2 poob Kev sim chloroquine yog ib qho kev sim classic rau kev soj ntsuam autophagy flux. Txhawm rau nkag siab txog cov laj thawj ntawm kev nce hauv LC3-II thiab P62 tom qab Sidt2 tshem tawm, peb siv chloroquine (CQ), cov dej ntws qis ntawm autophagy, ua rau cov hlwb los soj ntsuam cov autophagy flux. Ua ntej, peb siv cov concentration gradients sib txawv kom paub meej tias qhov saturation concentration inhibited los ntawm CQ yog 50 μM (Fig. 4A–F) hauv MPC5 thiab SV40 MES 13 hlwb. Nyob rau sab saum toj ntawm qhov concentration, thaum CQ concentration txuas ntxiv mus, LC3-II thiab P62 tsis tau nce ntxiv kom ncav cuag saturation. Yog li ntawd, peb siv 50 μM CQ stimulation dhau 16 h kom tag nrho inhibit autophagy flux, thiab lub sijhawm no, LC3-II qhov sib txawv ntawm Sidt2+/+ pab pawg thiab Sidt2-/- pab pawg ploj hauv MPC5 thiab SV40 MES 13 hlwb. Ib yam li ntawd, tom qab 50 μM CQ ua tiav, P62 qhov sib txawv tshwm sim los ntawm kev tshem tawm ntawm Sidt2 kuj ploj (Daim duab 4G, H, J, K). Cov txheeb cais ntawm autophagy flux tau pom tias nws txo qis thaum Sidt2 ploj lawm (Daim duab 4I, L), uas tau lees paub ntxiv tias qhov nce hauv LC3-II thiab P62 qhia tom qab Sidt2 tshem tawm yog vim tsis ua tiav ntawm autophagy clearance. es tsis yog qhov nce qib hauv autophagy (kev ua kom autophagy).
Kev cuam tshuam ntawm autophagosome-lysosome fusion hauv vitro tom qab Sidt2 deletion Clearance thiab degradation yog nruab nrab thiab lig theem nyob rau hauv cov txheej txheem autophagy, nrog rau fusion ntawm autophagosome nrog txo tom qab deletion ntawm Sidt2 (Fig. 5D), qhia tias fusion ntawm autophagosomes thiab lysosomes raug cuam tshuam.
Daim duab 1Lub raum puasthiab autophagolysosome txuam nrog hauv Sidt2-/- nas. Cre-LoxP system gene targeting schematic; B saum toj no yog daim duab qhia kev xa tawm ntawm Sidt2 noob knockout nas tail DNA; Hauv qab no yog daim ntawv qhia ua ntu zus, lub xub qhia qhov chaw ntawm cov noob uas ploj lawm. Piv nrog WT nas, 199 bp noob poob tshwm sim hauv exon 2; C DNA theem pov thawj ntawm Sidt2 (tshuaj los ntawm cov ntaub so ntswg). Cov khoom lag luam primer amplified muaj lub hauv paus knockout cheeb tsam, pom li Sidt2+ /+(WT), Sidt2+/−, los yog Sidt2−/−; D protein qib pov thawj kom pom Sidt2 protein qhia qib los ntawm sab hnub poob blot; E raum 24 h zis protein nyob rau hauv WT thiab Sidt2−/− nas; F ultra-micro-morphological qauv ntawm ob lub raum ntawm WT nas (a, f) thiab Sidt2−/− nas (g–n). Piv nrog rau WT nas, Sidt2-/- nas lub raum qhia txog cov txheej txheem ntawm ko taw fusion, hauv qab daus membrane thickening (g, h), raum tubular epithelial cell edema, microvilli puas (i, j), mitochondrial kev puas tsuaj (k, l), thiab autophagolysosome txuam nrog (m, n); G tag nrho cov raum autophagolysosomes hauv WT thiab Sidt2-/- nas. *P < 0 05, **P < 0.01.
Fig. 3 Sidt2 deletion cuam tshuam txoj kev autophagy. Kev Tshawb Nrhiav WT thiab Sidt2−/- cell autophagy-related protein qhia qib los ntawm sab hnub poob blot; B statistical charts of the western blot test results; C P62 mRNA qhia theem hauv raum cov ntaub so ntswg ntawm WT thiab Sidt2−/− nas; D qhia theem ntawm MPC5 cell autophagy pathway proteins ua ntej thiab tom qab Sidt2 knockout; E statistical graph ntawm (D) daim ntawv; F qhia theem ntawm autophagy pathway proteins hauv SV40 MES 13 hlwb ua ntej thiab tom qab Sidt2 knockout; G daim ntawv txheeb cais ntawm daim ntawv (F); H P62 immunofluorescence hauv MPC5 thiab SV40 MES 13 hlwb ua ntej thiab tom qab Sidt2 knockout; Kuv daim ntawv txheeb cais ntawm cov txiaj ntsig immunofluorescence; J P62 mRNA qhia theem hauv MPC5 thiab SV40 MES 13 hlwb ua ntej thiab tom qab Sidt2 knockout. *P <0.05, **P <0.01, ***P <0.001.
Ad-mCherry-GFP-
LC3B fluorescence ob-labeling qhia tias autophagolysosome tsim thiab nws txoj kev degradation raug thaiv tom qab Sidt2 deletion Ad-mCherry-GFP-LC3B transfection sim siv tau los kuaj xyuas cov kev hloov pauv hauv autophagy flux hauv hlwb. Thaum lub sij hawm tus txheej txheem ntawm autophagy, mCherry-GFP-LC3B sib sau ua ke ntawm autophagosome membrane thiab manifests nws tus kheej nyob rau hauv daim ntawv ntawm daj me ntsis nyob rau hauv ib tug fluorescence microscope. Thaum autophagosomes thiab lysosomes raug fused los ua autophagolysosomes, acidic ib puag ncig nyob rau hauv lub lysosome yuav quench lub fluorescence ntawm GFP kom nws manifests nws tus kheej nyob rau hauv daim ntawv ntawm liab me ntsis. Yog li ntawd, GFP khi rau LC3 tsuas yog siv los kuaj autophagosomes, thaum mCherry tuaj yeem ntes autophagosomes thiab autophagolysosomes tib lub sijhawm. Thaum cov fluorescence ntsuab thiab liab sib xyaw ua ke thiab tso tawm raws li cov xim daj fluorescence, qhov no sib raug rau autophagosomes. Lub sijhawm no, cov fluorescence liab tuaj yeem qhia tau tias yog autophagolysosomes, thiab nws tuaj yeem qhia qhov smoothness ntawm autophagolysosome tsim [27]. MPC5 hlwb thiab SV40 MES 13 hlwb nyob rau hauv ob lub Sidt2+/+ thiab Sidt2−/− pab pawg tau kis nrog Ad-mCherry GFP-LC3B adenovirus thiab yees duab nrog confocal laser microscope (Fig. 6A). Nws tau pom tias cov dots daj fluorescent tau nce thiab cov liab fluorescent dots txo qis (Daim duab 6B, C) tom qab Sidt2 tshem tawm, pom tias cov autophagosomes nce thiab cov autophagolysosomes txo qis hauv pawg Sidt2-/-. Qhov no yog vim muaj kev cuam tshuam rau kev tsim autophagolysosome, thiab txoj hauv kev autophagolysosome degradation raug thaiv, uas ua rau autophagosome degradation obstacles.
Rapamycin tsis tau txhim kho P62 tsub zuj zuj los ntawm Sidt2 tshem tawm tab sis nws ua rau nws hnyav dua Rapamycin (RAPA) yog ib qho inhibitor ntawm mTOR thiab activates autophagy. Thaum RAPA ua rau MPC5 thiab SV40 MES 13 hlwb ntawm Sidt2+/+ thiab Sidt2−/− pab pawg, nws nce LC3-II hauv ob pawg. Qhov no txhais tau hais tias RAPA tuaj yeem qhib txoj hauv kev ntawm autophagy. Ib yam li ntawd, peb pom tias qhov nce hauv LC3-II theem hauv Sidt2−/- pab pawg nrog RAPA tau pom tseeb dua li hauv kev tswj hwm (Sidt2+/+ pawg), qhia tias thaum RAPA ua ntawm Sidt2-/- pab pawg, qhov degradation ntawm autophagosomes tseem tsis tau txhim kho. Lub caij no, peb kuj tau soj ntsuam cov kev hloov pauv hauv P62 thiab pom tias thaum RAPA ua rau Sidt2+/+ pab pawg MPC5 thiab SV40 MES 13 hlwb, P62 tsis hloov pauv, qhia tau hais tias autophagy flux yog qhov qub. Thaum RAPA ua rau pawg Sidt2−/−, kev qhia theem ntawm P62 tau nce ntxiv piv rau qhov ua ntej kev tswj hwm thiab tseem pom tseeb dua li kev tswj hwm tom qab kev kho RAPA (Fig. 7A, B, D, E). Tom qab ntawd, txhawm rau tshawb nrhiav cov laj thawj ntawm qhov tsis sib xws ntawm P62 protein tom qab Sidt2+/+ thiab Sidt2-/- pab pawg tau kho nrog RAPA, peb ntsuas mRNA qib ntawm P62 thiab pom tias qib nce ntxiv. Tom qab RAPA kev kho mob hauv ob pawg Sidt2+/+ thiab Sidt2−/− pawg, thiab Sidt2+/+ pab pawg muaj qib siab dua li hauv pawg Sidt2−/− (Fig. 7C , F), uas tuaj yeem ua pov thawj ntxiv tias qhov autophagy ntws tau zoo nyob hauv Sidt 2+/+ pab pawg, thaum lub autophagy flow disorder tshwm sim thaum kawg ntawm autophagy hauv pawg Sidt2-/-, uas yog, autophagolysosomes degradation txuas. Peb kuj tau kawm txog cov txiaj ntsig ntawm Sidt2 ntawm kev loj hlob thiab apoptosis ntawm lub raum hlwb nyob rau hauv cov xwm txheej ntawm activated autophagy thiab pom tias tom qab cov ntshav tsis muaj qhov nruab nrab ntawm cov cell ua kom autophagy, piv nrog pawg tswj hwm, kev loj hlob ntawm Sidt2-/-. pab pawg tau inhibited ntxiv, thiab qhov apoptosis tau pom tseeb dua (Cov duab ntxiv 2).
Fig. 5 Autophagosome thiab lysosome fusion yog tiv thaiv tom qab Sidt2 deletion. A Immunofluorescence co-localization ntawm LC3B thiab LAMP1 hauv MPC5 thiab SV40 MES 13 hlwb ua ntej thiab tom qab Sidt2 knockout, tsom xam nrog Pearson qhov sib txheeb coefficient; B sib piv ntawm LC3B fluorescence cov ntsiab lus ua ntej thiab tom qab Sidt2 knockout hauv MPC5 thiab SV40 MES 13 hlwb; C sib piv ntawm Pearson qhov sib txheeb coefficient ua ntej thiab tom qab Sidt2 knockout hauv MPC5 thiab SV40 MES 13 hlwb; D kev sib piv ntawm fusion tus nqi ua ntej thiab tom qab Sidt2 knockout hauv MPC5 thiab SV40 MES 13 hlwb (qhov piv ntawm tus naj npawb ntawm cov chaw sib koom ua ke fluorescent me ntsis ntawm LAMP1 thiab LC3B rau cov naj npawb ntawm fluorescent me ntsis ntawm LAMP1); *P <0.05, **P <0.01.
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