beta actin 8h10d10  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Rab12 is a regulator of mitophagy and mitochondrial homeostasis

doi: 10.64898/2026.03.29.715103

Figure Lengend Snippet: ( A ) RNA was collected for qRT-PCR analysis of Rab12 mRNA expression in mt-mKeima/YFP-parkin HeLa cells following 72 h transfection with siRNA against Rab12 (siRab12) or the scramble control (siScr). Internal control for normalization was GAPDH. **** p < 0.0001 determined by unpaired t-test. n = 3 biological replicates with three technical replicates each. ( B ) Representative western blot of lysates collected from mt-mKeima/YFP-parkin HeLa cells transfected with siScr or siRab12 assessed for Rab12 levels and β-actin as a loading control. ( C ) Quantification demonstrates Rab12 protein knockdown with Rab12 siRNA by the same protocol. **** p < 0.0001, as determined by unpaired t-test. n = 3 biological replicates. ( D ) Representative 60X confocal images of YFP-Parkin (green), mt-mKeima at neutral pH (cyan), and mt-mKeima at lysosomal pH (magenta) and ( E ) quantification of mt-mKeima mitophagy analysis demonstrated increased levels of mitophagy induced by treatment with 30 μM FCCP with Rab12 knockdown. ** p < 0.01, **** p < 0.0001, as determined by two-way ANOVA with Bonferroni’s multiple comparisons test. n = 3 biological replicates. Data are presented as mean ± SEM.

Article Snippet: For our analysis, we utilized the following antibodies: rabbit anti-LRRK2 (MJFF2 c41-2), (Abcam, ab133474, 1:2000), sheep anti-Rab12 (MRC-PPU, SA227, 0.5 μg/mL), and mouse β-actin (Novus Biologicals, NBP1-47423, 1:10,000).

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

Journal: Molecules

Article Title: Epacadostat and Olaparib Synergistically Inhibit the Growth of BRCA-Proficient Triple-Negative Breast Cancer by Suppressing the Expression of BRCA1 and RAD51

doi: 10.3390/molecules31061039

Figure Lengend Snippet: The effects of Epacadostat and/or Olaparib on the expression of BRCA1 and RAD51 in vitro. Cells were pre-stimulated with IFN-γ (50 ng/mL) for 24 h to induce the over-expression of IDO1 before treatment. ( A ) Quantitative PCR analysis of BRCA1 and RAD51 mRNA levels following the indicated treatments. Data were normalized to TBP and expressed as fold change relative to the control group. ( B ) Western blot analysis of BRCA1 and RAD51 proteins in the tumor cells treated with Epacadostat and/or Olaparib. ( C ) The protein levels BRCA1 was quantified by ImageJ (ImageJ, RRID: SCR_003070) and normalized to the β-tubulin protein content. ( D ) The protein levels RAD51 was quantified by ImageJ and normalized to the β-tubulin protein content. “+” means the compound was added and “-” means the compound was not added. All data are presented as the mean ± SD of three independent experiments. Statistical significance was determined using one-way ANOVA. * p < 0.05 and ** p < 0.01 compared with control group, ## p < 0.01 compared with the combination group of Epacadostat and Olaparib.

Article Snippet: BRCA1 Antibody (1:1000, #9010), RAD51 (D4B10) Rabbit mAb (1:1000, #8875), anti-mouse IgG-HRP-linked antibody (1:3000, #7076), anti-rabbit IgG -HRP-linked antibody (1:3000, #7074), and anti-β-tubulin (8H10D10) mouse mAb (1:1000, #3700) were purchased from Cell Signaling Technology (Boston, MA, USA).

Techniques: Expressing, In Vitro, Over Expression, Real-time Polymerase Chain Reaction, Control, Western Blot

Journal: Molecules

Article Title: Epacadostat and Olaparib Synergistically Inhibit the Growth of BRCA-Proficient Triple-Negative Breast Cancer by Suppressing the Expression of BRCA1 and RAD51

doi: 10.3390/molecules31061039

Figure Lengend Snippet: The effect of Epacadostat and/or Olaparib on the expression of BRCA1 and RAD51 in vivo. ( A ) Quantitative PCR analysis of BRCA1 and RAD51 mRNA levels following the indicated treatments. Data were normalized to GAPDH and expressed as fold change relative to the control group. ( B ) Western blot analysis of BRCA1 and RAD51 proteins in the tumor tissues of mice treated with Epacadostat and/or Olaparib. ( C ) The protein levels of BRCA1 were quantified by ImageJ and normalized to the β-tubulin protein content. ( D ) The protein levels of RAD51 were quantified by ImageJ and normalized to the β-tubulin protein content. All data are presented as the mean ± SD of three independent experiments. Statistical significance was determined using one-way ANOVA. * p < 0.05 and ** p < 0.01 compared with control group, ## p < 0.01 compared with the combination group of Epacadostat and Olaparib.

Article Snippet: BRCA1 Antibody (1:1000, #9010), RAD51 (D4B10) Rabbit mAb (1:1000, #8875), anti-mouse IgG-HRP-linked antibody (1:3000, #7076), anti-rabbit IgG -HRP-linked antibody (1:3000, #7074), and anti-β-tubulin (8H10D10) mouse mAb (1:1000, #3700) were purchased from Cell Signaling Technology (Boston, MA, USA).

Techniques: Expressing, In Vivo, Real-time Polymerase Chain Reaction, Control, Western Blot