Journal: Annals of Translational Medicine
Article Title: Blocking MAPK/ERK pathway sensitizes hepatocellular carcinoma cells to temozolomide via downregulating MGMT expression
doi: 10.21037/atm-20-5478
Figure Lengend Snippet: U0126 enhanced TMZ-induced DNA damage. (A) Protein expression levels of p-ATR, γ-H2AX, p-ATM, p-ERK1/2 and ERK1/2 were detected when HepG2 cells were treated with TMZ (1 mM) and U0126 (4 and 8 µM) for 24 h by Western blot analysis (*, P<0.05, **, P<0.01 vs. the indicated treatment). TMZ, temozolomide. (B) Immunofluorescence staining analysis of HepG2 cells for γ-H2AX (red). DAPI (blue) was used to stain the nuclei (representative fields at ×200 magnification). TMZ, temozolomide; MAPK/ERK, mitogen-activated protein kinase/extracellular signal regulated kinase pathway; MGMT, methylguanine-DNA methyltransferase.
Article Snippet: The filters were blocked using 5% nonfat milk (Mengniu, Beijing, China) at room temperature for 2 h. Primary antibodies against MGMT (CST; 1:2,000), p-ATR (CST; 1:1,000), p-ATM (CST; 1:1,000), γ-H2AX (CST; 1:1,000), and β-actin (CST; 1:10,000) were added to filters and incubated overnight at 4 °C.
Techniques: Expressing, Western Blot, Immunofluorescence, Staining