anti beclin1  (Cell Signaling Technology Inc)

Journal: Veterinary Research

Article Title: Outer membrane vesicles secreted by avian pathogenic Escherichia coli promote its survival within macrophages and systemic infection by inducing endoplasmic reticulum stress-mediated autophagy flux blockade

doi: 10.1186/s13567-025-01679-6

Figure Lengend Snippet: APEC OMVs induce incomplete autophagy in HD11 cells . A HD11 cells were treated with OMVs (100 µg/mL) for 6 h, and typical double-membrane autophagosomes were observed by transmission electron microscopy. Scale bar,500 nm. B , C HD11 cells were treated with OMVs (100 µg/mL) for 0–9 h, western blot analysis was performed to detect the expression of LC3-II, p62, and Beclin1 proteins in cell lysates, and gray value analysis was performed using ImageJ software ( C ) ( n = 3). D Immunofluorescence was used to detect the effect of OMVs (100 µg/mL) on the colocalization of LC3 and the lysosomal marker protein LAMP1 in HD11 cells. The data are representative of three independent experiments. Scale bar, 5 μm. Colocalization scatter plots were generated using ImageJ software and GraphPad Prism 8. n represents three biological replicates, with each data point corresponding to an independent culture system. Bar charts display mean ± standard error of the mean (SEM), analyzed by Student’s -test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Rabbit anti-Beclin1 polyclonal antibody , Bioss, Beijing, China , 1:2000.

Techniques: Membrane, Transmission Assay, Electron Microscopy, Western Blot, Expressing, Software, Immunofluorescence, Marker, Generated

Journal: Antioxidants

Article Title: Epicatechin Gallate Ameliorates UVB-Induced Photoaging by Inhibiting p38α-Mediated Autophagy and Oxidative Stress

doi: 10.3390/antiox15020180

Figure Lengend Snippet: Effects of ECG on autophagy in HaCaT cells. ( A ) Representative immunofluorescence images of autophagic vacuoles stained with MDC (scale bar: 200 μm) and quantitative analysis of MDC fluorescence intensity. ( B ) Representative Western blots and densitometric analysis of LC3B, p62, and Beclin1 proteins. ( C ) Relative mRNA expression levels of BECN1 , MAP1LC3B , and SQSTM1/p62 determined by RT-qPCR. ( D ) Representative transmission electron microscopy images of HaCaT cells (scale bar: 500 nm). Red arrows indicate autophagosome-like structures; lowercase letters ‘m’ denotes mitochondria. ( E ) Representative Western blots and densitometric analysis of LC3B and p62 proteins. Data are presented as the mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 and ns denotes not significant.

Article Snippet: Rabbit anti-Beclin1 (BECN1) antibody (cat. #11306), mouse anti-tubulin antibody (cat. #66031-1-IG), rabbit anti-LaminB1 (LMNB1) antibody (cat. #12987-1-AP), rabbit anti-Cyclin-Dependent Kinase Inhibitor 1A (p21, CDKN1A) antibody (cat. #10355-1-AP) and rabbit anti-LC3 antibody (cat. #14600) were purchased from Proteintech group (Wuhan, China).

Techniques: Immunofluorescence, Staining, Fluorescence, Western Blot, Expressing, Quantitative RT-PCR, Transmission Assay, Electron Microscopy

Journal: Antioxidants

Article Title: Epicatechin Gallate Ameliorates UVB-Induced Photoaging by Inhibiting p38α-Mediated Autophagy and Oxidative Stress

doi: 10.3390/antiox15020180

Figure Lengend Snippet: Role of p38α inhibition in ECG-mediated effects on autophagy and ROS. ( A ) Representative Western blots and densitometric analysis of Beclin1, p62, and LC3B proteins in HaCaT cells treated with ECG and/or a p38α inhibitor. ( B ) Relative mRNA expression levels of BECN1, MAP1LC3B , and SQSTM1/p62 under the same treatments as in ( A ). ( C ) Representative immunofluorescence images of intracellular ROS levels (scale bar: 100 μm) and quantitative analysis. ( D ) Representative immunofluorescence images of MDC staining (scale bar: 200 μm) and quantitative analysis under the same treatments. ( E ) Representative images of SA-β-gal staining (scale bar: 200 μm) and quantitative analysis under the same treatments. Data are presented as the mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: Rabbit anti-Beclin1 (BECN1) antibody (cat. #11306), mouse anti-tubulin antibody (cat. #66031-1-IG), rabbit anti-LaminB1 (LMNB1) antibody (cat. #12987-1-AP), rabbit anti-Cyclin-Dependent Kinase Inhibitor 1A (p21, CDKN1A) antibody (cat. #10355-1-AP) and rabbit anti-LC3 antibody (cat. #14600) were purchased from Proteintech group (Wuhan, China).

Techniques: Inhibition, Western Blot, Expressing, Immunofluorescence, Staining

Journal: Antioxidants

Article Title: Epicatechin Gallate Ameliorates UVB-Induced Photoaging by Inhibiting p38α-Mediated Autophagy and Oxidative Stress

doi: 10.3390/antiox15020180

Figure Lengend Snippet: Effects of ECG in p38α-overexpressing HaCaT cells. ( A ) Schematic of the experimental timeline for p38α overexpression and ECG treatment. ( B ) Validation of p38α overexpression by RT-qPCR and Western blot analysis of p38α and p-p38α. ( C – F ) Representative Western blots and densitometric analysis of Beclin1, p62, and LC3B proteins in control and p38α-overexpressing cells with or without ECG treatment. ( G – I ) Activities of SOD, CAT, and GSH-Px in cell lysates under the indicated conditions. ( J ) Flow cytometric analysis of intracellular ROS levels under the indicated conditions. ( K , M ) Representative immunofluorescence images of γH2AX and Lamin B1 (scale bar: 100 μm) under the indicated conditions. White arrows indicate nuclei with damage. ( L , N ) Representative images of PI incorporation assay (scale bar: 100 μm) and quantitative analysis of PI-positive cells. ( O , P ) Representative images of SA-β-gal staining (scale bar: 200 μm) and quantitative analysis of positive cells. Data are presented as the mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: Rabbit anti-Beclin1 (BECN1) antibody (cat. #11306), mouse anti-tubulin antibody (cat. #66031-1-IG), rabbit anti-LaminB1 (LMNB1) antibody (cat. #12987-1-AP), rabbit anti-Cyclin-Dependent Kinase Inhibitor 1A (p21, CDKN1A) antibody (cat. #10355-1-AP) and rabbit anti-LC3 antibody (cat. #14600) were purchased from Proteintech group (Wuhan, China).

Techniques: Over Expression, Biomarker Discovery, Quantitative RT-PCR, Western Blot, Control, Immunofluorescence, Staining

Journal: European Journal of Medical Research

Article Title: SIN3A promotes lung adenocarcinoma by repressing MIR22HG/Beclin1 axis-mediated autophagy and ferroptosis

doi: 10.1186/s40001-026-03852-9

Figure Lengend Snippet: MIR22HG activates both autophagy and ferroptosis by mediating Beclin1 in LUAD. A Relative mRNA expression levels of Beclin1 in H1975 and H1299 cells transfected with control siRNA (siCtrl) or two independent siRNAs targeting Beclin1 (siBeclin1-1 and siBeclin1-2), as determined by RT-qPCR. B Immunoblot analysis of Beclin1, p62, LC3B, SLC7A11, and GPX4 in H1975 and H1299 cells following Beclin1 knockdown, with or without CQ (10 μM, 24 h) treatment. GAPDH was used as a loading control. C Immunoblot analysis of the same proteins in H1299 cells transduced with control lentivirus (LV-Ctrl) or MIR22HG-overexpressing lentivirus (LV-MIR22HG), combined with siRNA-mediated Beclin1 knockdown and/or CQ (10 μM, 24 h) treatment. D ROS levels in H1975 and H1299 cells measured by flow cytometry following Beclin1 gene silencing. E RIP experiments investigating the regulatory relationship between MIR22HG and Beclin1

Article Snippet: LUAD cells were lysed to obtain total protein extracts, and equal amounts of protein (approximately 20 μg) were separated by 12% SDS–PAGE at 80 V for 40 min followed by 120 V for 50 min, then transferred onto PVDF membranes at 90 V for 50 min. Membranes were blocked with 3% BSA for 30 min and incubated overnight at 4 °C with primary antibodies against Beclin1 (Cell Signaling Technology, Danvers, MA, USA; Cat# 3495, 1:1000), LC3B (Cat# 3868, 1:1000), p62/SQSTM1 (Cat# 5114, 1:1000), SLC7A11 (Cat# 12691, 1:1000), GPX4 (Cat# 52455, 1:1000), and SIN3A (Cat# 12872, 1:1000).

Techniques: Expressing, Transfection, Control, Quantitative RT-PCR, Western Blot, Knockdown, Transduction, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Pterostilbene attenuates osteoarthritis progression through p53-dependent autophagy activation: evidence from network analysis and experimental validation

doi: 10.3389/fphar.2026.1686555

Figure Lengend Snippet: PT protects chondrocytes against IL-1β-induced damage via autophagy activation. (A) Representative WB images showing ACAN, COL2A1, MMP3, MMP13, Beclin1, LC3B, and p62 expression; (B–H) Quantitative analysis of protein levels normalized to GAPDH for ACAN (B) , COL2A1 (C) , MMP13 (D) , MMP3 (E) , Beclin1 (F) , LC3II/I ratio (G) , and p62 (H) ; (I) Representative TUNEL staining showing apoptotic cells (red) and nuclei (blue) (scale bar = 50 μm); (J) Percentage of TUNEL-positive cells. Data = mean ± SEM (n = 3). **P < 0.01, ***P < 0.001 vs. control; ###P < 0.001 vs. IL-1β; †P < 0.05, †††P < 0.001 vs. IL-1β + PT.

Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) in TBST for 1 h and incubated overnight at 4 °C with the following primary antibodies: LC3B (cells 1:1000, tissue 1:800; #3868, Cell Signaling Technology(CST), Danvers, MA, United States), Beclin1 (cells 1:1000, tissue 1:800; #3495, CST), p62 (cells 1:1000, tissue 1:800; #88588, CST), collagen type II alpha 1 chain (COL2A1; cells 1:1000, tissue 1:500; #ab34712, Abcam, Cambridge, UK), aggrecan (ACAN; cells 1:500, tissue 1:500; #sc-33695, Santa Cruz Biotechnology, Dallas, TX, United States), matrix metalloproteinase 3 (MMP3; cells 1:1000, tissue 1:500; #14351, CST), matrix metalloproteinase 13 (MMP13; cells 1:1000, tissue 1:500; #69926, CST), p-AMPKα (Thr172; 1:1000; #2535, CST), AMPKα (1:1000; #2532, CST), p-mTOR (Ser2448; 1:1000; #5536, CST), mTOR (1:1000; #2983, CST), p53 (1:1000; #2527, CST), PCNA (1:2000; #13110, CST), and GAPDH (cells 1:10,000, tissue 1:5000; #5174, CST).

Techniques: Activation Assay, Expressing, TUNEL Assay, Staining, Control

Journal: Frontiers in Pharmacology

Article Title: Pterostilbene attenuates osteoarthritis progression through p53-dependent autophagy activation: evidence from network analysis and experimental validation

doi: 10.3389/fphar.2026.1686555

Figure Lengend Snippet: PT regulates cartilage-specific and autophagy-related protein expression in MIA-induced OA in vivo . (A) Representative WB images showing ACAN, COL2A1, MMP3, MMP13, Beclin1, LC3B, and p62 expression; (B–H) Quantitative analysis of protein levels normalized to GAPDH for ACAN (B) , COL2A1 (C) , MMP3 (D) , MMP13 (E) , Beclin1 (F) , LC3II/I ratio (G) , and p62 (H) . Data = mean ± SEM (n = 3). ###P < 0.001 vs. sham; *P < 0.01, **P < 0.001 vs. MIA; †P < 0.05, ††P < 0.01, †††P < 0.001 vs. PT-L.

Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) in TBST for 1 h and incubated overnight at 4 °C with the following primary antibodies: LC3B (cells 1:1000, tissue 1:800; #3868, Cell Signaling Technology(CST), Danvers, MA, United States), Beclin1 (cells 1:1000, tissue 1:800; #3495, CST), p62 (cells 1:1000, tissue 1:800; #88588, CST), collagen type II alpha 1 chain (COL2A1; cells 1:1000, tissue 1:500; #ab34712, Abcam, Cambridge, UK), aggrecan (ACAN; cells 1:500, tissue 1:500; #sc-33695, Santa Cruz Biotechnology, Dallas, TX, United States), matrix metalloproteinase 3 (MMP3; cells 1:1000, tissue 1:500; #14351, CST), matrix metalloproteinase 13 (MMP13; cells 1:1000, tissue 1:500; #69926, CST), p-AMPKα (Thr172; 1:1000; #2535, CST), AMPKα (1:1000; #2532, CST), p-mTOR (Ser2448; 1:1000; #5536, CST), mTOR (1:1000; #2983, CST), p53 (1:1000; #2527, CST), PCNA (1:2000; #13110, CST), and GAPDH (cells 1:10,000, tissue 1:5000; #5174, CST).

Techniques: Expressing, In Vivo