Journal: Molecular Therapy. Nucleic Acids

Article Title: Combination of microRNA-21 and microRNA-146a Attenuates Cardiac Dysfunction and Apoptosis During Acute Myocardial Infarction in Mice

doi: 10.1038/mtna.2016.12

Figure Lengend Snippet: Changes of protein levels of PTEN, TRAF6, p-AKT, and p-p38 in AMI mice. ( a ) Representative western blot bands of PTEN protein. n = 5/group. ( b ) Representative western blot bands of TRAF6 protein. n = 4/group. ( c ) Representative western blot bands of p-AKT and AKT proteins. n = 4/group. ( d ) Representative western blot bands of p-p38 and p38 proteins. Values given were normalized to the band intensity of β-actin as an internal control. n = 4/group (* P < 0.05, ** P < 0.01, one-way analysis of variance).

Article Snippet: PTEN siRNA and TRAF6 siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Western Blot, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Combination of microRNA-21 and microRNA-146a Attenuates Cardiac Dysfunction and Apoptosis During Acute Myocardial Infarction in Mice

doi: 10.1038/mtna.2016.12

Figure Lengend Snippet: Synergistic suppression of p-p38 MAPK by the miR-21: miR-146a pair in neonatal rat ventricular myocytes . ( a,b ) Verification of the efficacy and specificity of TRAF6 siRNA in silencing TRAF6 at both mRNA and protein levels. n = 4/group. ( c ) Effect of TRAF6 siRNA on the protein level of p-p38. n = 4/group. ( d,e ) Verification of the efficacy and specificity of PTEN siRNA in silencing PTEN at both mRNA and protein levels. n = 4/group. ( f,g ) Changes of p-p38 and p-AKT levels in PTEN siRNA transfected cells. n = 4/group. ( h,i ) Effects of PI3K inhibitor LY294002 on p-AKT and p-p38 levels. Cells were preincubated with LY294002 (10 μmol/l) for 2 hours, then transfected with miR-21 for 36 hours, and finally treated with hypoxia for 12 hours. n = 4/group (* P < 0.05, ** P < 0.01, one-way analysis of variance). ( j ) The diagram illustrating the synergistic mechanism of miR-21: miR-146a pair attenuates apoptosis.

Article Snippet: PTEN siRNA and TRAF6 siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Transfection

Journal: Molecules (Basel, Switzerland)

Article Title: Berberine Inhibited Growth and Migration of Human Colon Cancer Cell Lines by Increasing Phosphatase and Tensin and Inhibiting Aquaporins 1, 3 and 5 Expressions.

doi: 10.3390/molecules28093823

Figure Lengend Snippet: Figure 11. Effect of berberine on the expression of PTEN, AKT, p-AKT, mTOR and p-mTOR in CRC

Article Snippet: Membranes were subsequently incubated overnight with the following primary antibodies at 4 ◦C: PTEN (AF847, R&D System; 1:1000), p-AKT-S473 (AF887, R&D System; 1:1000), AKT (MAB2055, R&D System; 1:1000), p-mTOR-S2448 (ab109268, Abcam; 1:1000), mTOR (AHO1232, Thermo Fisher Scientific; 1:1000) and beta actin (2118S; Cell Signaling Technology, Inc.; Danvers, MA, USA; 1:1000).

Techniques: Expressing

Journal: Autophagy

Article Title: ATM-mediated PTEN phosphorylation promotes PTEN nuclear translocation and autophagy in response to DNA-damaging agents in cancer cells.

doi: 10.1080/15548627.2015.1009767

Figure Lengend Snippet: Figure 1. ATM regulated TPT-induced PTEN nuclear translocation in A549 and HeLa cells. (A) Repre- sentative photomicrographs showed endogenous PTEN translocation (100£). A549 and HeLa cells were treated with 3 mg/mL TPT and/or 10 mM KU55933 for 24 h. (B) 3£Flag-PTEN-WT plasmids were transfected into the HeLa cells, and the transfection efficiency was confirmed by western blot analysis. (C) Representative photomicrograph images showed exogenous PTEN translocation (100£). HeLa cells were transiently transfected with 3£Flag-PTEN-WT plasmids, and 24 h after transfection, the cells were treated with 3 mg/mL TPT and/or 10 mM KU55933 and/or ATM siRNA for 24 h. (D) Repre- sentative photomicrograph images showing exogenous PTEN translocation (100£). HeLa cells were transiently transfected with 3£Flag-PTEN-WT or 3£Flag-PTENS113A plasmids, and 24 h after transfec- tion, the cells were treated with 6 mg/mL CDDP. (E) Western blot analysis result of exogenous PTEN nuclear translocation in A549 and HeLa cells when treated with 3 mg/mL TPT and/or 10 mM KU55933 for 24 h. (F) Western blotting result of exogenous PTEN nuclear translocation in A549 and HeLa cells when treated with 3 mg/mL TPT and/or ATM siRNA for 24 h.

Article Snippet: Purified ATM protein (active) (14–933) was purchased from Millipore, and purified PTEN recombinant protein (TP710023) was obtained from Origene.

Techniques: Translocation Assay, Transfection, Western Blot

Journal: Autophagy

Article Title: ATM-mediated PTEN phosphorylation promotes PTEN nuclear translocation and autophagy in response to DNA-damaging agents in cancer cells.

doi: 10.1080/15548627.2015.1009767

Figure Lengend Snippet: Figure 2. ATM mediated TPT-induced PTEN phosphorylation. (A) Sequence analysis of PTEN was per- formed by Scansite software. The sequence of PTEN was retrieved from GenBank. (B) A549 cells were treated with 3 mg/mL TPT, l-phosphatase, and PPase inhibitors as indicated. Tagged proteins were analyzed by immunoblotting. (C) HeLa cells were transiently transfected with the 3£Flag-PTEN-WT plasmid，then treated with various concentrations of TPT or CDDP. (D) A549 and HeLa cells were transiently transfected with 3£Flag-PTEN-WT plasmids, and 24 h after transfection, the cells were treated with 3 mg/mL TPT; then, immunoprecipitation was conducted with an antibody against Flag. The precipitates were subjected to SDS-PAGE and probed with antibody against phospho-PTENS113. (E) A549 cells were treated with 3 mg/mL TPT for 24 h in the absence or presence of ATM siRNA- or KU55933, and then immunoblotting analysis was performed. (F) In vitro phosphorylation assays with active ATM kinase and purified PTEN recombinant proteins. Reaction products were subjected to SDS-PAGE and probed with antibody against phospho-PTENS113.

Article Snippet: Purified ATM protein (active) (14–933) was purchased from Millipore, and purified PTEN recombinant protein (TP710023) was obtained from Origene.

Techniques: Phospho-proteomics, Sequencing, Software, Western Blot, Transfection, Plasmid Preparation, Immunoprecipitation, SDS Page, In Vitro, Recombinant

Journal: Autophagy

Article Title: ATM-mediated PTEN phosphorylation promotes PTEN nuclear translocation and autophagy in response to DNA-damaging agents in cancer cells.

doi: 10.1080/15548627.2015.1009767

Figure Lengend Snippet: Figure 3. TPT-induced PTEN nuclear translocation was dependent on PTEN (S113) phosphorylation in A549 and HeLa cells. A549 cells and/or HeLa cells were transiently transfected with the 3£Flag-PTEN-WT or 3£Flag-PTENS113A plasmid, and 24 h after transfection, the cells were treated with or without 3 mg/mL TPT. (A) Representative photomicro- graph images showing PTEN translocation (200£). (B) The expression levels of PTEN and p-PTENS113 were analyzed by immunoblotting. (C) Representative photomicrograph images showing PTEN translocation (200£) after treatment of 10 ng/mL of leptomycin B for 24 h.

Article Snippet: Purified ATM protein (active) (14–933) was purchased from Millipore, and purified PTEN recombinant protein (TP710023) was obtained from Origene.

Techniques: Translocation Assay, Phospho-proteomics, Transfection, Plasmid Preparation, Expressing, Western Blot

Journal: Autophagy

Article Title: ATM-mediated PTEN phosphorylation promotes PTEN nuclear translocation and autophagy in response to DNA-damaging agents in cancer cells.

doi: 10.1080/15548627.2015.1009767

Figure Lengend Snippet: Figure 4. The PTENS113A mutant enhanced TPT-induced DNA damage in A549 and HeLa cells. (A) A549 and HeLa cells were transiently transfected with the 3£Flag-PTEN-WT or 3£Flag-PTENS113A plasmid, and 24 h after transfection, the cells were treated with or without 3 mg/mL TPT. Representative photo- micrograph images showed accumulation of g-H2AFX as an indicator of DNA damage (100£). (B) Bar graphs representing the percentage of g-H2AFX-positive cells upon treatment with TPT for 24 h (*P < 0.05, **P <0.001, t test, n D 3 , bars represent SEM, Cells containing more than 5 foci were scored as positive). (C) A549 and HeLa cells were transiently transfected with the 3£Flag-PTEN-WT or 3£Flag- PTENS113A plasmid, and 24 h after transfection, the ells were treated with or without 3 mg/mL TPT. The expression levels of g-H2AFX protein were analyzed by immunoblotting.

Article Snippet: Purified ATM protein (active) (14–933) was purchased from Millipore, and purified PTEN recombinant protein (TP710023) was obtained from Origene.

Techniques: Mutagenesis, Transfection, Plasmid Preparation, Expressing, Western Blot

Journal: Autophagy

Article Title: ATM-mediated PTEN phosphorylation promotes PTEN nuclear translocation and autophagy in response to DNA-damaging agents in cancer cells.

doi: 10.1080/15548627.2015.1009767

Figure Lengend Snippet: Figure 6. Effect of PTEN expression level on TPT-induced autophagy in A549 and HeLa cells. (A) A549 and HeLa cells were stably transfected with PTEN shRNA plasmids. Cells were treated with or without 3 mg/mL TPT for 24 h. The expression levels of PTEN, MAP1LC3/LC3, and SQSTM1/p62 protein were analyzed by immunoblotting. Bar graphs represent the relative MAP1LC3/LC3-II and SQSTM1/p62 protein levels normalized to that of GAPDH of different groups (*P < 0.05, **P < 0.001, t test, n D 3 ). (B) A549 and HeLa cells were transiently transfected with the 3£Flag-PTEN-WT plasmids. Cells were treated with or without 3mg/mL TPT for 24 h. The expression levels of Flag, PTEN, MAP1LC3/LC3, and SQSTM1/p62 protein were analyzed by immunoblotting. Bar graphs represent the relative MAP1LC3/LC3-II and SQSTM1/p62 protein levels normalized to that of GAPDH of different groups. (*P < 0.05, **P < 0.001, t test, n D 3 ). (C) The effect of PTENS113 phosphorylation on autophagy in A549 and HeLa cells. The expression levels of MAP1LC3/LC3 pro- tein were analyzed by immunoblotting. (D) HeLa cells were transiently transfected with the 3£Flag-PTEN-WT or 3£Flag-PTENS113A plasmid and YFP-LC3 plasmids. Twenty four h after transfection, the cells were treated with or without 3 mg/mL TPT. Green YFP autofluorescence (YFP-LC3) was analyzed by confocal microscopy (100£). Bar graphs represent the percentage of YFP- LC3-positive cells in control or TPT-treated group (*P < 0.05, **P < 0.001, t test, n D 3 , bars represent SEM, Cells containing more than 5 foci were scored as positive). (E) A549 and HeLa cells were treated with or without 3 mg/ mL TPT for 24 h, followed by treatment with 100 mM bafilomycin A1 (Baf.A1) as indicated. The expression levels of MAP1LC3/LC3 and SQSTM1/p62 protein were analyzed by immunoblotting.

Article Snippet: Purified ATM protein (active) (14–933) was purchased from Millipore, and purified PTEN recombinant protein (TP710023) was obtained from Origene.

Techniques: Expressing, Stable Transfection, Transfection, shRNA, Western Blot, Phospho-proteomics, Plasmid Preparation, Confocal Microscopy, Control

Journal: Autophagy

Article Title: ATM-mediated PTEN phosphorylation promotes PTEN nuclear translocation and autophagy in response to DNA-damaging agents in cancer cells.

doi: 10.1080/15548627.2015.1009767

Figure Lengend Snippet: Figure 7. PTEN promoted TPT-induced autophagy via activation of the p-JUN-SESN2-AMPK pathway in A549 and HeLa cells. (A) The cells were treated with or without 3 mg/mL TPT, and the expression levels of JUN, p-JUN, SESN2, AMPK, p-AMPK, RPS6KB, and p-RPS6KB protein were analyzed by immunoblot- ting. (B) The cells were treated with or without 3 mg/mL TPT and/or JUN siRNA for 24 h, and the expres- sion levels of JUN, p-JUN, SESN2, AMPK, p-AMPK, RPS6KB, p-RPS6KB, and LC3 protein were analyzed by immunoblotting. Bar graphs represent the relative MAP1LC3/LC3-II protein levels normalized to that of GAPDH of different groups. (*P < 0.05, **P < 0.001, t test, n D 3 ). (C) The cells were treated with or with- out 3 mg/mL TPT and/or AMPK siRNA for 24 h, and the expression levels of p-AMPK, AMPK, RPS6KB, p-RPS6KB, and MAP1LC3/LC3 protein were analyzed by immunoblotting. Bar graphs represent the rel- ative MAP1LC3/LC3-II protein levels normalized to that of GAPDH of different groups. (*P <0.05, **P < 0.001, t test, n D 3 ). (D) The cells were treated with or without 3 mg/mL TPT, and the expression levels of p-GSK3B, GSK3B, p-AKT, and AKT protein were analyzed by immunoblotting.

Article Snippet: Purified ATM protein (active) (14–933) was purchased from Millipore, and purified PTEN recombinant protein (TP710023) was obtained from Origene.

Techniques: Activation Assay, Expressing, Western Blot

Journal: Life

Article Title: Spermine Ameliorates DSS-Induced Ulcerative Colitis in Mice by Improving Mitophagy and Intestinal Microbiota

doi: 10.3390/life16030417

Figure Lengend Snippet: Spermine treatment inhibited NLRP3-mediated inflammatory response in DSS-treated mice. ( A – E ) Genes expressions related to NLRP3-mediated inflammatory response in the colon were measured. ( F – I ) The levels of pro-inflammatory cytokines in the serum. ( J ) Detection of NLRP3 inflammasome protein expression levels in the colon using immunohistochemistry. Data are presented as mean ± SEM (* p < 0.05, *** p < 0.001 vs. control group. ## p < 0.01, and ### p < 0.001 vs. DSS group).

Article Snippet: The sections were then incubated at 4 °C overnight with primary antibodies targeting NLRP3 (1:500, Proteintech, 30109-1-AP, Chicago, IL, USA), PINK1 (1:500, Proteintech, 23274-1-AP, Chicago, IL, USA), and LC3 (1:500, Proteintech, 14600-1-AP, Chicago, IL, USA).

Techniques: Expressing, Immunohistochemistry, Control

Journal: Life

Article Title: Spermine Ameliorates DSS-Induced Ulcerative Colitis in Mice by Improving Mitophagy and Intestinal Microbiota

doi: 10.3390/life16030417

Figure Lengend Snippet: Potential mechanism of Spermine treatment alleviating DSS-induced colitis. First, spermidine enhances mitochondrial autophagy by upregulating the expression of PINK1/Parkin, thereby eliminating damaged mitochondria and reducing the production of reactive oxygen species. This, in turn, inhibits the activation of the NLRP3 inflammasome and the subsequent release of IL-1β/IL-18, thereby alleviating the inflammatory response. Secondly, spermidine can reverse the intestinal flora imbalance induced by DSS, manifested as a decrease in the ratio of Firmicutes to Bacteroidetes, inhibition of Blautia enrichment, and restoration of the abundance of beneficial Muribaculaceae. These two pathways work together in synergy, jointly forming an integrated mechanism by which spermidine inhibits the NLRP3-mediated inflammatory response, promotes mitochondrial autophagy, and improves intestinal flora imbalance to alleviate DSS-induced colitis.

Article Snippet: The sections were then incubated at 4 °C overnight with primary antibodies targeting NLRP3 (1:500, Proteintech, 30109-1-AP, Chicago, IL, USA), PINK1 (1:500, Proteintech, 23274-1-AP, Chicago, IL, USA), and LC3 (1:500, Proteintech, 14600-1-AP, Chicago, IL, USA).

Techniques: Expressing, Activation Assay, Inhibition

Journal: Molecular Medicine

Article Title: Complement protein C1q induces endothelial apoptosis through C1qBP regulation of mitochondrial function in lung endothelial cells in systemic sclerosis

doi: 10.1186/s10020-026-01445-0

Figure Lengend Snippet: C1qBP affected PHB2 phosphorylation and cytoplasmic translocation as well as mitochondrial dysfunction in endothelial cells. BLM-induced endothelial cells were treated with si-C1qBP. A - B Western blot analysis was utilized to examine the effects of si-C1qBP on C1qBP levels and the phosphorylation and cytoplasmic translocation of PHB2. C - D The binding and co-localization of C1qBP with PHB2 were observed through co-immunoprecipitation and immunofluorescence. E - F Western blot was performed to evaluate the impact of C1qBP on the expression of apoptosis-related proteins (BAX and Bcl-2), mitophagy-related proteins (Beclin1, Parkin, Pink1), as well as UPRmt-related markers (mtDnaJ, ClpP, LonP1, Hsp10). G Red CMXRos staining was used to assess the effect of C1qBP on mitochondrial activity in endothelial cells. H The influence of C1qBP on intracellular Ca 2+ levels in endothelial cells was measured by Fluo-4 AM fluorescent probe detection. I - J MMP detection was conducted to observe the effect of C1qBP on the MMP of endothelial cells. K - L The impact of C1qBP on ATP levels and electron transport chain (complexes I-IV) activity in endothelial cells was determined by ATP colorimetric/fluorescence assay and mitochondrial electron transport chain activity detection. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Con/ BLM + si-NC

Article Snippet: Antibodies used included: C1qA Primary Antibody (1:1,000, A1821, Abclonal), C1qB Primary Antibody (1:1,000, BD-PB4657, Biodragon), C1qBP Primary Antibody (1:1,000, 68084-1-Ig, Proteintech), COV IV Primary Antibody (1:1, 000, 4850 S, Cellular), PHB2 primary antibody (1:1,000, 66424-1-Ig, Proteintech), Cleaved caspase-3 primary antibody (1:1,000, 68773-1-Ig, Proteintech), ICAM1 primary antibody (1:1,000, A20472, Abclonal), ZO1 primary antibody (1:1,000, 21773-1-AP, Proteintech), ZO2 primary antibody (1:1,000, 18900-1-AP, Proteintech), OCLN primary antibody (1:1,000, 27260-1-AP, Proteintech), JAM3 primary antibody (1:1,000, BD-PN0240, Biodragon), BAX primary antibody (1:1,000, CY5059, Abways), Bcl-2 primary antibody (1:1,000, ab182858, Abcam), Beclin1 primary antibody (1:1,000, 3738 S, Cell signalling), Parkin primary antibody (1:1,000, CY6641, Abways), Pink1 primary antibody (1:1,000, BY0130, Abways), ClpP primary antibody (1:1,000, ab124822, Abcam), mtDnaJ primary antibody (1:1,000, ab124822, Abcam), LonP1 primary antibody (1:1,000, A03808-2, BOSTER), Hsp10 primary antibody (1:1,000, CY8473, Abways), and β-actin primary antibody (1:1,000, 66009-1-Ig, Proteintech).

Techniques: Phospho-proteomics, Translocation Assay, Western Blot, Binding Assay, Immunoprecipitation, Immunofluorescence, Expressing, Staining, Activity Assay, Fluorescence

Journal: Molecular Medicine

Article Title: Complement protein C1q induces endothelial apoptosis through C1qBP regulation of mitochondrial function in lung endothelial cells in systemic sclerosis

doi: 10.1186/s10020-026-01445-0

Figure Lengend Snippet: C1qBP promoted endothelial cell apoptosis by causing mitochondrial dysfunction through affecting the cytosolic translocation of PHB2. A PHB2-S293A phosphorylation-deficient mutant was constructed. BLM-induced endothelial cells were co-transfected with PHB2-S293A (WT or MUT) and siRNA (si-NC or si-C1qBP), respectively. A Western blot analysis of C1qBP, PHB2 and p-PHB2 expression. B Western blot analysis of apoptosis- and mitophagy-related proteins BAX, Bcl-2, Beclin1, Parkin, and Pink1. C - D Immunofluorescence detection of BAX, Bcl-2, Beclin1, Parkin and Pink1 expression. E Western blot analysis of mitochondrial unfolded protein response (UPRmt)-related markers mtDnaJ, ClpP, LonP1 and Hsp10. F Red CMXRos staining to detect mitochondrial activity. G Fluo-4 AM fluorescent probe to measure intracellular Ca 2+ levels. H Evaluation of mitochondrial electron transport chain (ETC) activity. I JC-1 assay to detect mitochondrial membrane potential (MMP). J ATP colorimetric/fluorometric assay kit to measure ATP levels. K Flow cytometry analysis of the number of viable endothelial cells. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. BLM + si-NC+PHB2-S293A WT/ BLM + si-C1qBP+PHB2-S293A WT/ BLM + si-NC+PHB2-S293A MUT. ns means no significant difference vs. BLM + si-NC+PHB2-S293A WT/ BLM + si-C1qBP+PHB2-S293A WT

Article Snippet: Antibodies used included: C1qA Primary Antibody (1:1,000, A1821, Abclonal), C1qB Primary Antibody (1:1,000, BD-PB4657, Biodragon), C1qBP Primary Antibody (1:1,000, 68084-1-Ig, Proteintech), COV IV Primary Antibody (1:1, 000, 4850 S, Cellular), PHB2 primary antibody (1:1,000, 66424-1-Ig, Proteintech), Cleaved caspase-3 primary antibody (1:1,000, 68773-1-Ig, Proteintech), ICAM1 primary antibody (1:1,000, A20472, Abclonal), ZO1 primary antibody (1:1,000, 21773-1-AP, Proteintech), ZO2 primary antibody (1:1,000, 18900-1-AP, Proteintech), OCLN primary antibody (1:1,000, 27260-1-AP, Proteintech), JAM3 primary antibody (1:1,000, BD-PN0240, Biodragon), BAX primary antibody (1:1,000, CY5059, Abways), Bcl-2 primary antibody (1:1,000, ab182858, Abcam), Beclin1 primary antibody (1:1,000, 3738 S, Cell signalling), Parkin primary antibody (1:1,000, CY6641, Abways), Pink1 primary antibody (1:1,000, BY0130, Abways), ClpP primary antibody (1:1,000, ab124822, Abcam), mtDnaJ primary antibody (1:1,000, ab124822, Abcam), LonP1 primary antibody (1:1,000, A03808-2, BOSTER), Hsp10 primary antibody (1:1,000, CY8473, Abways), and β-actin primary antibody (1:1,000, 66009-1-Ig, Proteintech).

Techniques: Translocation Assay, Phospho-proteomics, Mutagenesis, Construct, Transfection, Western Blot, Expressing, Immunofluorescence, Staining, Activity Assay, Membrane, ATP Colorimetric Fluorometric Assay, Flow Cytometry

Journal: Molecular Medicine

Article Title: Complement protein C1q induces endothelial apoptosis through C1qBP regulation of mitochondrial function in lung endothelial cells in systemic sclerosis

doi: 10.1186/s10020-026-01445-0

Figure Lengend Snippet: C1qBP knockdown promoted mitophagy, alleviated endothelial cell apoptosis, and pulmonary fibrosis. A - B Immunohistochemical detection of BAX, Bcl-2, Beclin1, Parkin, and Pink1 expression. C TUNEL and CD31 co-staining assay was conducted to detect apoptosis. D Evaluation of pulmonary microvascular permeability in mice by Evans Blue extravasation assay. E Co-localization of pimonidazole with CD31 in fibrotic lung tissue. F Immunofluorescence co-staining of CD31 with HIF-1α

Article Snippet: Antibodies used included: C1qA Primary Antibody (1:1,000, A1821, Abclonal), C1qB Primary Antibody (1:1,000, BD-PB4657, Biodragon), C1qBP Primary Antibody (1:1,000, 68084-1-Ig, Proteintech), COV IV Primary Antibody (1:1, 000, 4850 S, Cellular), PHB2 primary antibody (1:1,000, 66424-1-Ig, Proteintech), Cleaved caspase-3 primary antibody (1:1,000, 68773-1-Ig, Proteintech), ICAM1 primary antibody (1:1,000, A20472, Abclonal), ZO1 primary antibody (1:1,000, 21773-1-AP, Proteintech), ZO2 primary antibody (1:1,000, 18900-1-AP, Proteintech), OCLN primary antibody (1:1,000, 27260-1-AP, Proteintech), JAM3 primary antibody (1:1,000, BD-PN0240, Biodragon), BAX primary antibody (1:1,000, CY5059, Abways), Bcl-2 primary antibody (1:1,000, ab182858, Abcam), Beclin1 primary antibody (1:1,000, 3738 S, Cell signalling), Parkin primary antibody (1:1,000, CY6641, Abways), Pink1 primary antibody (1:1,000, BY0130, Abways), ClpP primary antibody (1:1,000, ab124822, Abcam), mtDnaJ primary antibody (1:1,000, ab124822, Abcam), LonP1 primary antibody (1:1,000, A03808-2, BOSTER), Hsp10 primary antibody (1:1,000, CY8473, Abways), and β-actin primary antibody (1:1,000, 66009-1-Ig, Proteintech).

Techniques: Knockdown, Immunohistochemical staining, Expressing, TUNEL Assay, Staining, Permeability, Immunofluorescence