Supplementary Figure 5: Telomerase activity modulates sensitivity to MTH1 depletion

(a) Immunoblot with antibodies against MTH1 or actin. The percent MTH1 expression relative to the controls are shown. (b) Cells were immuno-stained with an 8oxoG antibody (green) 3 days after infection with lentivirus expressing a non-targeting shRNA (scr) or two different shRNAs targeting MTH1 (sh4 or sh5). Nuclei are indicated by DAPI staining (blue). Exposure times and image settings were equivalent for the sh4 and sh5, compared to the scr control. (c) BJhTERT and BJ cells were fixed 3 days after lentiviral infection and stained for SA-beta-gal activity. The percentage of positive cells is indicated on the bar graph. (d) Whole cell lysates (10μg) were immunoblotted with anti-p53 and anti-actin antibody as a loading control 3 days after lentiviral infection. (e) MTH1 knock down impairs BJ-hTERT proliferation as compared to BJ fibroblasts. DIC images were captured 12 days following lentiviral infection with a 10x objective.