rabbit anti human bcl2  (Proteintech)

Journal: World Journal of Diabetes

Article Title: Dl-3-n-butylphthalide ameliorates diabetic foot ulcer by inhibiting apoptosis and promoting angiogenesis

doi: 10.4239/wjd.v16.i4.101916

Figure Lengend Snippet: Molecular docking analysis and mRNA expression analysis of core genes. A: Molecular docking revealed the binding of Dl-3-n-butylphthalide to its targets. Blue solid line: Hydrogen bond; Gray dashed line: Hydrophobic interaction; Yellow dashed line: Electrostatic interaction; B: The quantitative real-time polymerase chain reaction analysis of the mRNA levels of heme oxygenase 1, caspase 3, B cell leukemia/lymphoma 2, brain derived neurotrophic factor and nuclear factor erythroid 2 L2. Data expressed as individual values with mean ± SD. b P < 0.01, a P < 0.05. HMOX: Heme oxygenase; CASP3: Caspase 3; BCL2: B cell leukemia/lymphoma 2; BDNF: Brain derived neurotrophic factor; NFE2: Nuclear factor erythroid 2; HG: High glucose; NBP: Dl-3-n-butylphthalide.

Article Snippet: The primary antibodies used were rabbit anti-human B cell leukemia/lymphoma 2 (BCL2, Proteintech, Wuhan, China, 1:2000), rabbit anti-human BCL2 associated X (Proteintech, Wuhan, China, 1:2000), rabbit anti-human advanced glycosylation end-product specific receptor (Proteintech, Wuhan, China, 1:3000), and mouse anti-human β-actin (Proteintech, Wuhan, China, 1:5000).

Techniques: Expressing, Binding Assay, Real-time Polymerase Chain Reaction, Derivative Assay

Journal: Cancer Research

Article Title: Targeting BCL2 with Venetoclax Enhances the Efficacy of the KRAS G12D Inhibitor MRTX1133 in Pancreatic Cancer

doi: 10.1158/0008-5472.CAN-23-3574

Figure Lengend Snippet: BCL2 inhibitor venetoclax enhances the efficacy of MRTX1133 in “high”-confluent collagen cultures. A, PDAC cells (2138, 3213, 1245, and PANC1) in “high”-confluent collagen cultures were treated with MRTX1133 (0.5 µmol/L) or the MEK1/2 inhibitor trametinib (0.1 µmol/L) for 8 hours. The effect on BIM and BCL2 was analyzed by Western blotting. Blots are representative of at least three biological replicates. B, PDAC cells in “high”-confluent collagen cultures were cotreated with venetoclax (2.5 µmol/L) and MRTX1133 (0.5 µmol/L) for 8 hours, and the effect on cell death (c-C3, cleaved-PARP) was analyzed. Blots are representative of at least three biological replicates. C, “High”-confluent collagen cultures of PDAC cells were cotreated with venetoclax (V; 2.5 µmol/L) and MRTX1133 (M; 0.5 µmol/L) for 48 hours, and the effect on cell growth was analyzed. Error bars, ± SD; n = 3 to 4. One-way ANOVA, followed by the Tukey multiple comparison test. D, PDAC cells were transfected with control siRNA or siRNA against BIM for 72 hours, plated in collagen, and treated with MRTX1133 (0.5 µmol/L) for 8 hours. The effect on BIM was analyzed using Western blotting. The transfected cells were also cotreated with venetoclax (2.5 µmol/L) and MRTX1133 (0.5 µmol/L) for 8 hours, and the effect on cell death (c-C3) was analyzed. Blots are representative of three biological replicates. E, “High”-confluent collagen cultures of PDAC cells were cotreated with venetoclax (2.5 µmol/L) and MRTX1133 (0.5 µmol/L) for 48 hours with or without the pan-caspase inhibitor zVAD-FMK (zVAD; 10 µmol/L), and the effect on cell growth was analyzed. Error bars, ± SD, n = 3. One-way ANOVA, followed by the Tukey multiple comparison test. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: The following antibodies were used at the dilution recommended by the manufacturers: pERK1/2 (#9101, Cell Signaling Technology, RRID:AB_331646), total ERK1/2 (#9102, Cell Signaling Technology, RRID:AB_330744), mouse BIM (#2933, Cell Signaling Technology, RRID:AB_1030947), mouse BCL2 (554218, BD Pharmingen, RRID:AB_395311), cleaved caspase-3 (c-C3; #9664, Cell Signaling Technology, RRID:AB_2070042), human BCL2 (#3498, Cell Signaling Technology, RRID: AB_1903907), PARP (#9542, Cell Signaling Technology, RRID:AB_2160739), HSP90 (#4877, Cell Signaling Technology, RRID:AB_2233307), and GAPDH (MAB374, MilliporeSigma, RRID:AB_2107445).

Techniques: Western Blot, Comparison, Transfection, Control

Journal: Oncology Letters

Article Title: ZBTB20 suppresses tumor growth in glioblastoma through activating the TET1/FAS/caspase‑3 pathway

doi: 10.3892/ol.2024.14491

Figure Lengend Snippet: ZBTB20 promotes apoptosis in glioblastoma cells. (A) Western blotting was used to detect BCL2, BAX, CASP3 and cleaved CASP3 expression in ZBTB20-overexpressing U251 cells, (B) the levels of which were semi-quantified and are presented as histograms. The independent experiment was repeated three times. Data are presented as the mean ± standard deviation. *P<0.05. BCL2, B-cell lymphoma-2; BAX, BCL2 associated X; CASP3, caspase-3; NC, negative control; ZBTB20, zinc finger and BTB domain containing 20.

Article Snippet: Next, the membranes were separately incubated overnight at 4°C with the following antibodies: Polyclonal rabbit anti-human ZBTB20 (dilution, 1:2,000; cat. no. ab127702; Abcam), polyclonal rabbit anti-TET1 (1:1,000; cat. no. A21766; ABclonal Biotech Co., Ltd.), polyclonal rabbit anti-human FAS (1:1,000; cat. no. A2639; ABclonal Biotech Co., Ltd.), polyclonal rabbit anti-human B-cell lymphoma-2 (BCL2; 1:2,000; cat. no. BA0412; Wuhan Boster Biological Technology, Ltd.), polyclonal rabbit anti-human BCL2 associated X (BAX; 1:2,000, cat. no. BA0315-2; Wuhan Boster Biological Technology, Ltd.), monoclonal rabbit anti-human CASP3 (1:1,000, cat. no. AF1213; Beyotime Institute of Biotechnology), monoclonal rabbit anti-human cleaved CASP3 (1:1,000; cat. no. AF1150; Beyotime Institute of Biotechnology), monoclonal rabbit anti-human phosphorylated-extracellular signal regulated kinase (p-ERK; 1:1,000; cat. no. AP0485; ABclonal Biotech Co., Ltd.), polyclonal rabbit anti-human ERK (1:1,000; cat. no. A16686; ABclonal Biotech Co., Ltd.), monoclonal rabbit anti-human p-cyclic AMP-responsive-element-binding protein (CREB; 1:500; cat. no. ab32096; Abcam), monoclonal mouse anti-human CREB (1:500; cat. no. ab178322; Abcam) and monoclonal rabbit anti-human β-actin (1:10,000; cat. no. AC026; ABclonal Biotech Co., Ltd.).

Techniques: Western Blot, Expressing, Standard Deviation, Negative Control