Journal: International journal of molecular sciences

Article Title: Short-Term Starvation Weakens the Efficacy of Cell Cycle Specific Chemotherapy Drugs through G1 Arrest.

doi: 10.3390/ijms24032498

Figure Lengend Snippet: Figure 1. Short-term starvation protects cancer cells against ETO by reducing DNA damage. (A) Dose- response curves with IC50 values that indicate cell lines pre-starved or left untreated for 24 h were treated with the indicated concentration of ETO for 48 h. (B) Immunoblot (left) and quantification (right) of γH2AX in HeLa cells pre-starved or untreated for 24 h with 20 µM ETO treatment for 2 h following different recovery times. (C) Immunoblot (left) and quantification (right) of γH2AX in H460 cells pre-starved or untreated for 24 h with 20 µM ETO treatment for 2 h following different recovery times. (D) Comet assay images (left) showing the tail moment (right) of pre-starved or untreated HeLa cells with or without 20 µM ETO treatment for 2 h. Scale bar, 100 µm. Data is represented as mean ± SD of at least three independent experiments. p values are from student’s t-tests. ** p < 0.01; *** p < 0.001.

Article Snippet: The cells were harvested at various time points post-ETO and processed for the comet assay using a DNA Damage Detection Kit (SCGE) (KeyGEN BioTECH, Cat# KGA240) according to the manufacturer’s protocol.

Techniques: Concentration Assay, Western Blot, Single Cell Gel Electrophoresis

Journal: International journal of molecular sciences

Article Title: Short-Term Starvation Weakens the Efficacy of Cell Cycle Specific Chemotherapy Drugs through G1 Arrest.

doi: 10.3390/ijms24032498

Figure Lengend Snippet: Figure 3. GAPDH deficiency protects cancer cells is not attributed to increased DNA damage repair. (A) Immunofluorescence images (left) of γH2AX in HeLa cells transfected with scramble siRNA or si-GAPDH after 20 µM ETO treatment for 2 h. DNA was stained with DAPI (blue). Quantification (right) of γH2AX foci/cell. Scale bar, 10 µm. (B) Immunofluorescence images (left) of 53BP1 in HeLa cells transfected with scramble siRNA or si-GAPDH after 20 µM ETO treatment for 2 h. DNA was stained with DAPI (blue). Quantification (right) of 53BP1 foci/cell. Scale bar, 10 µm. (C) Immunoblot (left) and quantification (right) of γH2AX in control or GAPDH knockdown HeLa cells with 20 µM ETO treatment for different recovery times. (D) Comet assay images (left) showing the tail moment (right) of control and GAPDH knockdown HeLa cells with or without 20 µM ETO treatment. Scale bar, 100 µm. (E) Immunoblot (upper) and quantification (lower) of indicated proteins in HeLa cells transfected with an empty vector and Flag-GAPDH were treated with or without 20 µM ETO. (F) The relative HR efficiency of U2OS cells were transfected with scramble siRNA and siGAPDH. (G) The relative NHEJ efficiency of U2OS cells was transfected with scramble siRNA and siGAPDH. (H) Immunoblot (upper) and quantitative analysis (lower) of γH2AX levels in HeLa cells transfected with scrambled siRNA and si-GAPDH for 24 h were treated with NU7441 and RI-1 for 2 h, and then treated with ETO following different recovery times. Data represented as mean ± SD of at least three independent experiments. p values are from student’s t-tests. ** p < 0.01; *** p < 0.001.

Article Snippet: The cells were harvested at various time points post-ETO and processed for the comet assay using a DNA Damage Detection Kit (SCGE) (KeyGEN BioTECH, Cat# KGA240) according to the manufacturer’s protocol.

Techniques: Transfection, Staining, Western Blot, Control, Knockdown, Single Cell Gel Electrophoresis, Plasmid Preparation