rabbit anti beclin1 becn1 antibody  (Proteintech)

Journal: Animals : an Open Access Journal from MDPI

Article Title: Caffeic Acid Counteracts LPS-Induced Inflammatory Damage in Yak Mammary Epithelial Cells Associated with NF-κB-Mediated Autophagy Regulation

doi: 10.3390/ani16111605

Figure Lengend Snippet: CA alleviates LPS-induced inflammatory response. Cells were pretreated with different concentrations of CA (75, 150, and 300 μM) for 54 h in the absence or presence of LPS (5 μg/mL) for 48 h. ( A ) Immunofluorescence result for NF-κB, Scale bar = 10 μm. ( B – D ) Western blot analysis of Nrf2, NF-κB, and pNF-κB protein. ( E ) Immunofluorescence results for LC3, Scale bar = 10 μm. ( F , J , K ) Western blot analysis of LC3, NF-κB, and p-NF-κB protein. ( G – I ) Western blot analysis of Beclin-1, p62 protein. One-way ANOVA, Dunnett’s post hoc test. Data are presented as the mean ± SEM. ns p > 0.05 (not significant), * p < 0.05, ** p < 0.01.

Article Snippet: The membranes were then blocked with 5% BSA and incubated with primary antibodies targeting the NF-κB (1:500, ER0815, HUABIO, Hangzhou, China), p-NF-κB (1:500; HA723223, HUABIO, Hangzhou, China), Nrf2 (1:1000; HA721432, HUABIO, Hangzhou, China), p62 (1:1000, A22025, Abclonal, Wuhan, China), LC3 (1:1000, A19665, Abclonal, Wuhan, China), and Beclin-1 (1:200, A17028, Abclonal, Wuhan, China). β-actin was used as an internal control to normalize of target protein expression levels.

Techniques: Immunofluorescence, Western Blot

Journal: Animals : an Open Access Journal from MDPI

Article Title: Caffeic Acid Counteracts LPS-Induced Inflammatory Damage in Yak Mammary Epithelial Cells Associated with NF-κB-Mediated Autophagy Regulation

doi: 10.3390/ani16111605

Figure Lengend Snippet: The effect of inhibiting the Nrf2 signalling pathway on alleviating the inflammatory response in primary YMECs induced by LPS. Cells were pretreated with 150 μM concentrations of CA for 6 h, followed by the addition of LPS (5 μg/mL) and ML385 for an additional 48 h. The total treatment duration was 54 h. ( A ) Immunofluorescence result for LC3, Scale bar = 10 μm. ( B – F ) Western blot analysis of Nrf2, Beclin-1, p62, and LC3 protein. One-way ANOVA, Dunnett’s post hoc test. Data are presented as the mean ± SEM. ns p > 0.05 (not significant), * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The membranes were then blocked with 5% BSA and incubated with primary antibodies targeting the NF-κB (1:500, ER0815, HUABIO, Hangzhou, China), p-NF-κB (1:500; HA723223, HUABIO, Hangzhou, China), Nrf2 (1:1000; HA721432, HUABIO, Hangzhou, China), p62 (1:1000, A22025, Abclonal, Wuhan, China), LC3 (1:1000, A19665, Abclonal, Wuhan, China), and Beclin-1 (1:200, A17028, Abclonal, Wuhan, China). β-actin was used as an internal control to normalize of target protein expression levels.

Techniques: Immunofluorescence, Western Blot

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: Cancers

Article Title: Cabozantinib Sensitizes NSCLC Cells to Radiation by Inducing Ferroptosis via STAT3/MCL1/BECN1/SLC7A11 Axis Suppression

doi: 10.3390/cancers17182950

Figure Lengend Snippet: Cabozantinib promotes ferroptosis through the MCL1/BECN1/SLC7A11 pathway. ( A ) Glutamate release and total GSH levels in H1299 cells treated with DMSO, 4 Gy RT, 6.25 μM Cabo, or the combination of RT and Cabo. ( B ) Immunoprecipitation (IP) of BECN1 in cells under the same conditions as ( A ), followed by immunoblotting for SLC7A11, MCL1, and BECN1. ( C ) A549 cells were transfected with control siRNA (siCtrl) or BECN1 siRNA (siBECN1), treated with DMSO or the combination of radiation and cabozantinib. Immunoblotting analysis confirms the BECN1 knockdown; meanwhile, glutamate release and GSH levels were measured. ( D , E ) Lipid ROS (C11-BODIPY) and MDA levels ( D ) and PI-positive cells ( E ) were measured under the same conditions as ( C ). ( F ) Clonogenic survival of A549 cells expressing shCtrl or shBECN1 treated with DMSO or the combination of radiation and cabozantinib. Data are presented for at least three independent experiments. *** p < 0.001. The orange peak denotes PI-negative viable cells, while the pink peak represents PI-positive non-viable/dead cells.

Article Snippet: For each sample, 1000 μg of total protein was incubated overnight at 4 °C with 1 μg of anti-BECN1 antibody (Cell Signaling Technology, #3495).

Techniques: Immunoprecipitation, Western Blot, Transfection, Control, Knockdown, Expressing

Journal: Cancers

Article Title: Cabozantinib Sensitizes NSCLC Cells to Radiation by Inducing Ferroptosis via STAT3/MCL1/BECN1/SLC7A11 Axis Suppression

doi: 10.3390/cancers17182950

Figure Lengend Snippet: Prognostic significance of BECN1 and SLC7A11 expression in radiotherapy-treated NSCLC patients. Kaplan–Meier survival curves of NSCLC patients (n = 65) who received radiotherapy stratified by BECN1 ( A ) and SLC7A11 ( B ) expression levels. The hazard ratios with 95% confidence intervals and log-rank P -values are indicated. ( C ) Proposed model of the MCL1/BECN1/SLC7A11 axis regulating ferroptosis in response to radiotherapy in NSCLC.

Article Snippet: For each sample, 1000 μg of total protein was incubated overnight at 4 °C with 1 μg of anti-BECN1 antibody (Cell Signaling Technology, #3495).

Techniques: Expressing