puromycin n acetyltransferase gene  (OriGene)

Journal: Food Science & Nutrition

Article Title: Exploring the Mechanisms of Total Saponins of Black Ginseng and Ginsenoside Rg3 Against Doxorubicin‐Induced Cardiotoxicity

doi: 10.1002/fsn3.71968

Figure Lengend Snippet: The potential function and pathways of TSF on DOX‐induced cardiotoxicity: (a) PPI network of 103 overlapping targets constructed based on the STRING database; (b) the PPI network map; (c) the molecular docking of HDAC8 with BRCA2.

Article Snippet: Antibodies for BRCA2, DRP1 and Bcl‐2 were sourced from Wanlei Biotechnology Co. Ltd. (Lot: T11052438, T09113028, T11201556), while β ‐actin and HRP‐labeled secondary antibodies were procured from Affinity Biosciences (Lot: T0022, S0001).

Techniques: Construct

Journal: Food Science & Nutrition

Article Title: Exploring the Mechanisms of Total Saponins of Black Ginseng and Ginsenoside Rg3 Against Doxorubicin‐Induced Cardiotoxicity

doi: 10.1002/fsn3.71968

Figure Lengend Snippet: TSF and Rg3 regulate HDAC8/BRCA2/DRP1 pathway inhibits DOX‐induced cardiomyocyte apoptosis. (a) Apoptosis was assessed using TUNEL staining (Scale bar = 50 μm). (b) The results of western blotting for each group. (c) The expression of Bcl‐2 for each group. (d) The expression of DRP1 for each group. (e) The expression of HDAC8 for each group. (f) The expression of BRCA2 for each group. (g) Immunofluorescence staining for observing the colocalization of HDAC8, BRCA2, and DRP1 (Scale bar = 1 mm). Data are expressed as mean ± standard from three individual experiments. Data analysis among groups was performed using ANOVA, with LSD for equal variances and Dunnett’s‐T3 for unequal variances, with significance levels of * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Antibodies for BRCA2, DRP1 and Bcl‐2 were sourced from Wanlei Biotechnology Co. Ltd. (Lot: T11052438, T09113028, T11201556), while β ‐actin and HRP‐labeled secondary antibodies were procured from Affinity Biosciences (Lot: T0022, S0001).

Techniques: TUNEL Assay, Staining, Western Blot, Expressing, Immunofluorescence

Journal: Food Science & Nutrition

Article Title: Exploring the Mechanisms of Total Saponins of Black Ginseng and Ginsenoside Rg3 Against Doxorubicin‐Induced Cardiotoxicity

doi: 10.1002/fsn3.71968

Figure Lengend Snippet: The mechanism diagram of TSF and Rg3 alleviating DOX‐induced myocardial injury by regulating the HDAC8/BRCA2/DRP1 pathway.

Article Snippet: Antibodies for BRCA2, DRP1 and Bcl‐2 were sourced from Wanlei Biotechnology Co. Ltd. (Lot: T11052438, T09113028, T11201556), while β ‐actin and HRP‐labeled secondary antibodies were procured from Affinity Biosciences (Lot: T0022, S0001).

Techniques:

Journal: PLOS One

Article Title: Differential sensitivity of MCPH1- and BRCA2-deficient cancer cells to PARP-1 inhibition

doi: 10.1371/journal.pone.0345514

Figure Lengend Snippet: (A) . U2OS or Hela cells were transfected with Control (siCON), MCPH1 (siMCPH1) or BRCA2 (siBRCA2) siRNA for 48h or 120h and protein expression analysed via Western blotting with the indicated antibodies. MCPH1 expression in sgCON (control) and sgMCPH1 (MCPH1 knockout) cells was also investigated by Western blotting. (B) Quantification of MCPH1 or BRCA2 protein bands normalised to GAPDH and expressed as % change relative to siCON or sgCON. Data is the mean and standard error of three (48h) or two (120h and sgCON/sgMCPH1) independent experiments. (C) U2OS or Hela cells were transfected with MCPH1 siRNA for 72h before analysis of nuclear morphology (premature chromosome condensation, PCC) via DAPI staining and fluorescence microscopy. (D) The % of cells displaying PCC, with at least 100 cells analysed.

Article Snippet: Membranes were blocked with 5% dried skimmed milk/TBS-T (50mM Tris pH 7.6, 150mM NaCl and 0.2% v/v Tween-20) for a minimum of 1-hour prior to overnight incubation at 4°C with primary antibodies; MCPH1 (11962–1-AP, Proteintech), BRCA2 (29450–1-AP, Proteintech) and GAPDH (60004–1-Ig, Proteintech).

Techniques: Transfection, Control, Expressing, Western Blot, Knock-Out, Staining, Fluorescence, Microscopy

Journal: PLOS One

Article Title: Differential sensitivity of MCPH1- and BRCA2-deficient cancer cells to PARP-1 inhibition

doi: 10.1371/journal.pone.0345514

Figure Lengend Snippet: U2OS cells were transfected with control (siCON), BRCA2 (siBRCA2) or BRCA2 + MCPH1 (siBRCA2 + siMCPH1) siRNA for 48h before (A) Western blot analysis or (B) seeding into 96-well plates and treating with the indicated doses of AZD2461 or Talazoparib, with each treatment performed in triplicate. After 96-hours, cell viability was measured using an MTS assay. MTS data represents the mean and standard error from at least three independent experiments, with cell viability expressed as a % relative to the corresponding untreated sample. Statistical significance was measured using two-way ANOVA (p-values described in text, with significance set at p < 0.05).

Article Snippet: Membranes were blocked with 5% dried skimmed milk/TBS-T (50mM Tris pH 7.6, 150mM NaCl and 0.2% v/v Tween-20) for a minimum of 1-hour prior to overnight incubation at 4°C with primary antibodies; MCPH1 (11962–1-AP, Proteintech), BRCA2 (29450–1-AP, Proteintech) and GAPDH (60004–1-Ig, Proteintech).

Techniques: Transfection, Control, Western Blot, MTS Assay