phospho sqstm1 p62  (Cell Signaling Technology Inc)

Journal: Frontiers in Pharmacology

Article Title: Baicalin chemosensitivity enhancement of cisplatin in bladder cancer via autophagy flux inhibition

doi: 10.3389/fphar.2026.1676788

Figure Lengend Snippet: Cisplatin induces autophagy in BC T24 and BIU-87 cells. (A,B) T24 and BIU-87 cells were analyed by western blotting using anti-LC3B and anti-p62 antibodies after treating with cisplatin (1 μg/ml) for 24 h (C-E) T24 and BIU-87 cells were treated with cisplatin (1 μg/ml) for 24 h before being labeled with immunofluorescence staining of LC3B (red). The nuclei were counterstained with DAPI (blue). Scale bar: 20 µm. (F) The subcellular structure of BC T24 and BIU-87 cells after treatment with or without cisplatin (1 μg/ml) for 24 h were imaged by TEM (The green triangle are autophagosome, while the red triangle are autolysosomes). Scale bar: 1 μm. All the data were presented as means ± S.D. and are representative of three independent experiments. **P < 0.01; ***P < 0.001.

Article Snippet: Then the sections were incubated with 0.3% H 2 O 2 for 10 min at room temperature, washed with PBS and blocked in 5% BSA at 37 °C for 30 min, followed by incubated with primary antibodies: LAMP1 (1:200, 21997, Proteintech), CTSD (1:500, ab75852, Abcam), LC3B (1:200, 83506, CST), Phospho-SQSTM1/p62 (1:200, 16177, CST) overnight at 4 °C and the secondary antibody for 1 h at room temperature.

Techniques: Western Blot, Labeling, Immunofluorescence, Staining

Journal: Frontiers in Pharmacology

Article Title: Baicalin chemosensitivity enhancement of cisplatin in bladder cancer via autophagy flux inhibition

doi: 10.3389/fphar.2026.1676788

Figure Lengend Snippet: Confocal images of BC T24 and BIU-87 cells infected with adenovirus mCherry-GFP-LC3B (mCherry: red, eGFP: green, Hoechst: bule). (A-C) Western blot analysis of the protein level of LC3B I, LC3B II and P62 in T24 and BIU-87 cells treated with DMSO or baicalin (40 μM, 80 μM) for 24 h (D,E) The relative mRNA expression levels of LAMP1, LAMP2, CTSB, and CTSD were detected in T24 cells and BIU-87 cells following treatment with DMSO (as a control) or baicalin (40 μM). (F-I) The cells were infected with adenovirus for 24 h and then incubated with DMSO, cisplatin (1 μg/mL), Rapa (2.5 μM), CQ (25 μM), and baicalin (40 μM) for 24 h. Nuclei were stained with Hoechst. Scale bar: 20 μm. All the data were presented as means ± S.D. and are representative of three independent experiments. **P < 0.01; ***P < 0.001.

Article Snippet: Then the sections were incubated with 0.3% H 2 O 2 for 10 min at room temperature, washed with PBS and blocked in 5% BSA at 37 °C for 30 min, followed by incubated with primary antibodies: LAMP1 (1:200, 21997, Proteintech), CTSD (1:500, ab75852, Abcam), LC3B (1:200, 83506, CST), Phospho-SQSTM1/p62 (1:200, 16177, CST) overnight at 4 °C and the secondary antibody for 1 h at room temperature.

Techniques: Infection, Western Blot, Expressing, Control, Incubation, Staining

Journal: Frontiers in Pharmacology

Article Title: Baicalin chemosensitivity enhancement of cisplatin in bladder cancer via autophagy flux inhibition

doi: 10.3389/fphar.2026.1676788

Figure Lengend Snippet: Baicalin blocked the autophagic flux of BC in vivo by inhibiting lysosome activity. (A-C) The expression of LC3B, p62, CTSB, CTSD, LAMP1, and LAMP2 of xenograft tumors was detected by western blotting. (D,E) Immunohistochemical staining was performed on tumor tissue using LC3B, P62, LAMP1, CTSB, and CTSD antibodies. Scale bar: 100 μm. All the data were presented as means ± S.D. and are representative of three independent experiments. *P < 0.05 vs . PBS ; **P < 0.01 vs . PBS ; ***P < 0.001 vs . PBS.

Article Snippet: Then the sections were incubated with 0.3% H 2 O 2 for 10 min at room temperature, washed with PBS and blocked in 5% BSA at 37 °C for 30 min, followed by incubated with primary antibodies: LAMP1 (1:200, 21997, Proteintech), CTSD (1:500, ab75852, Abcam), LC3B (1:200, 83506, CST), Phospho-SQSTM1/p62 (1:200, 16177, CST) overnight at 4 °C and the secondary antibody for 1 h at room temperature.

Techniques: In Vivo, Activity Assay, Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Experimental & Molecular Medicine

Article Title: Cancer-associated fibroblast-derived extracellular vesicles regulate lipophagy through PLIN2 to modulate dormancy in salivary gland adenoid cystic carcinoma cells

doi: 10.1038/s12276-025-01600-3

Figure Lengend Snippet: a , b Representative Nile Red staining images of dormant SACC cells overexpressing PLIN2 in the absence or presence of HCQ treatment. Scale bar, 20 μm. c qRT–PCR analysis of dormant SACC cells transfected with a NC or PLIN2-overexpressing vector to measure ER stress-related gene expression. d Western blot analysis of p-EIF2S1, EIF2S1, p-PERK and PERK levels in dormant SACC cells. e The dormant SACC cells were incubated with PERK inhibitor GSK2606414 (10 μM). Then, the protein levels were determined by western blot. f Molecular modeling analysis of the interaction between PLIN2 and the binding site within the p62 domain. g , h Co-IP of p62, followed by western blot for p62 and PLIN2, in dormant SACC-LM cells ( g ) and SACC-LM cells overexpressing PLIN2 ( h ).

Article Snippet: The following antibodies were used: anti-NR2F1 (ab181137) and anti-LC3 (ab192890) from Abcam; anti-Atg5 (12994), anti-p-mTOR (5536) and anti-p62 (16177) from Cell Signaling Technology; anti-caspase-3 (68773-1-Ig), anti-p-ATK (66444-1-Ig), anti-BAX (50599-2-Ig), anti-FAP (11779-1-AP), anti-BCL2 (12789-1-AP) and anti-α-SMA (14395-1-AP) from Proteintech; anti-HSP70 (ET1601-11), anti-GAPDH (ET1601-4), anti-CD63 (ET1607-2), anti-p21 (HA722685), anti-PLIN2 (ET1704-17), anti-p27 (ET1608-61) and anti-EIF2S1 ( HA500385 ) from HUABIO; anti-p-PERK (340846), anti-PERK ( R25311 ) from Zen-Bio.

Techniques: Staining, Quantitative RT-PCR, Transfection, Plasmid Preparation, Gene Expression, Western Blot, Incubation, Binding Assay, Co-Immunoprecipitation Assay

Journal: International Journal of Nanomedicine

Article Title: Honeysuckle-Derived Exosome-Like Nanovesicles Protect Against Acute Liver Failure by Modulating Gut Microbiota

doi: 10.2147/IJN.S526819

Figure Lengend Snippet: HNVs promote autophagy and inhibit apoptosis thereby reducing inflammatory response. ( A ) Western blotting results of p53, p62, Atg5, Atg7. ( B–D ) Expression of inflammatory factors IL-6, IL-1β and TNF-α in serum by ELISA (n=6 in each group). ( E–G ) Using quantitative real-time PCR to identify the liver’s expression of the inflammatory factors IL-6, IL-1β and TNF-α (n=6 in each group). ( H ) TUNEL staining fluorogram. ( I ) Quantitative results of TUNEL staining (n=6 in each group). Statistical analysis was performed by one-way ANOVA. Data were expressed as the mean ± SD. * P < 0.05, ** P <0.01, n.s., not significant vs Model group.

Article Snippet: The following primary antibodies were employed: Marker (RM19001, ABclonal), Beta Actin Polyclonal antibody (20536-1-AP, Proteintech), GAPDH (10494-1-AP, Proteintech), Atg7 (A19604, ABclonal), Atg5 (A19677, ABclonal), P53 (2524T, Cell Signaling Technology), P62 (16177T, Cell Signaling Technology).

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, TUNEL Assay, Staining

Journal: Cell Death Discovery

Article Title: Targeting ESR1 restores SQSTM1-dependent autophagy and sensitizes ER-positive breast cancer to oxidative and radiation stress

doi: 10.1038/s41420-025-02755-8

Figure Lengend Snippet: A Protein levels of Bcl-2, caspase 9 (full length/cleaved) and caspase 3 in T47D with or without ESR1 knockdown in a radiation dose-dependent manner. Actin was used as internal control. B Protein levels of COX2 and GPX4 in T47D with or without ESR1 knockdown in a radiation dose-dependent manner. Actin was used as internal control. C Protein levels of p-IRE1-α and IRE1-α in T47D with or without ESR1 knockdown in a radiation dose-dependent manner. Actin was used as internal control. D Protein levels of LC3B (I/II) and p62 in T47D isogenic model with or without CQ treatment. Actin was used as internal control. E Protein levels of LC3B (I/II) and p62 in T47D with or without ESR1 knockdown in a radiation dose-dependent manner. Actin was used as internal control. F Images of LC3B in ESR1 overexpression or knockdown model by immunofluorescence, respectively. DAPI was used as internal control. G Images of LAMP2 in ESR1 overexpression model by immunofluorescence. DAPI was used as internal control. The data from three independent experiments are presented as the means ± SEM.

Article Snippet: Phospho-p62 (Ser349) , 95697 , Cell signaling , 1:1000.

Techniques: Knockdown, Control, Over Expression, Immunofluorescence

Journal: Cell Death Discovery

Article Title: Targeting ESR1 restores SQSTM1-dependent autophagy and sensitizes ER-positive breast cancer to oxidative and radiation stress

doi: 10.1038/s41420-025-02755-8

Figure Lengend Snippet: A Protein levels of ATM, p-ATM (S1981), Ku70, Ku80, XRCC4, Rad51 and BRCA1 in T47D with or without ESR1 knockdown by H2O2 treatment. Actin was used as an internal control. B Images of TP53BP1 and γ-H2AX in T47D ESR1 knockdown model by H 2 O 2 treatment by immunofluorescence. Hoechst was used as an internal control. C Protein levels of ATM, p-ATM (S1981), Ku70, Ku80, XRCC4 and Rad51 in MCF7 with or without ESR1 overexpression by H 2 O 2 treatment. Actin was used as an internal control. D Images of TP53BP1 and γ-H2AX in MCF7 ESR1 overexpression model by H 2 O 2 treatment via immunofluorescence. Hoechst was used as an internal control. E Protein levels of LC3B (I/II) and p62 in T47D ESR1 knockdown model with or without CQ/ H 2 O 2 . Actin was used as an internal control. F Images of LC3B in ESR1 overexpression and knockdown model by H 2 O 2 treatment via immunofluorescence. Hoechst was used as an internal control. G Protein levels of LC3B (I/II) and p62 in T47D ESR1 knockdown model treated with H 2 O 2 with or without 3-MA. Actin was used as an internal control. H Images of LC3B in the MCF7 ESR1 overexpression model were taken after treatment with H₂O₂ for one hour and with or without 3-MA for three hours. Hoechst was used as the internal control. The data from three independent experiments are presented as the means ± SEM.

Article Snippet: Phospho-p62 (Ser349) , 95697 , Cell signaling , 1:1000.

Techniques: Knockdown, Control, Immunofluorescence, Over Expression

Journal: Cell Death Discovery

Article Title: Targeting ESR1 restores SQSTM1-dependent autophagy and sensitizes ER-positive breast cancer to oxidative and radiation stress

doi: 10.1038/s41420-025-02755-8

Figure Lengend Snippet: A Electrophoretic separation and Coomassie blue staining of MCF7 with and without ESR1 overexpression. B Venn diagram showing the overlapping molecules between ESR1 IP, vector IP and ESR1 with MG-132 treatment. C Top ranking of selected molecules from ESR1-based proteomics profiles. D Interaction affinity between p62 and ESR1 by pull-down assay in ESR1 overexpression model. E Protein levels of p62 in MCF7 ESR1 overexpression by MG-132 treatment. Actin was used as internal control. F Protein levels of phosphor-p62, p62 and ESR1 in MCF7 cells with and without E2 treatment. Actin was used as internal control. G Interaction affinity between p62 and ESR1 by pull-down assay after E2 treatment. H Survival fraction of ESR1 knockdown model with or without E2 treatment followed by irradiation. ( I ) Protein levels of LC3B and p62 in T-47D ESR1 isogenic model. Actin was used as internal control. ( J ) Images of TP53BP1 and γ-H2AX in T47D ESR1 knockdown model by E2 treatment combined with H2O2 by immunofluorescence. Hoechst was used as internal control. K Images of LC3B in T47D ESR1 knockdown model by E2 treatment, E2 combined with p62 overexpression and TamR via immunofluorescence. Hoechst was used as internal control. The data from three independent experiments are presented as the means ± SEM.

Article Snippet: Phospho-p62 (Ser349) , 95697 , Cell signaling , 1:1000.

Techniques: Staining, Over Expression, Plasmid Preparation, Pull Down Assay, Control, Knockdown, Irradiation, Immunofluorescence