FIGURE 2.
LSD1 expression and growth inhibition by NCL1 in embryonal carcinoma cells. (A) Western blotting of LSD1 in NTERA‐2 and Tera‐1 cells treated with DMSO control. (B) ChIP‐qPCR showing H3K9me2 enrichment at SOX2 and OCT4 promoter regions after NCL1 treatment of NTERA‐2 cells. (C) NCL1 dose–response curves for NTERA‐2 and Tera‐1 cells measured using the WST‐8 assay. (D) Cisplatin dose–response curves for parental NTERA‐2 and cisplatin‐adapted NTERA‐2R cells assessed by WST‐8 assay. The corresponding cisplatin IC₅₀ values were 0.53 and 3.18 μM, respectively (6.0‐fold increase in NTERA‐2R). (E) NCL1 dose–response curve for cisplatin‐adapted NTERA‐2R cells. (F) WST‐8 absorbance in parental NTERA‐2 cells treated with increasing concentrations of cisplatin alone or combined with 5 μM NCL1. Data in panels (C–F) are mean ± SD of replicate wells within the experiment displayed; error bars indicate SD. For panels (C, E, and F), one representative experiment is shown from three independent experiments with consistent qualitative trends. Statistical comparisons in these three panels were performed using replicate wells from the representative experiment, not the independent experiments as replicates. Symbols in panels (C, E) indicate differences from the corresponding untreated control; symbols in panel (F) indicate differences between cisplatin alone and cisplatin plus NCL1 at the indicated cisplatin concentrations. The indicated comparisons in panels (C, E, and F) were evaluated using multiple‐comparison testing. *p < 0.05; **p < 0.01; ****p < 0.0001. ChIP‐qPCR, chromatin immunoprecipitation quantitative PCR; LSD1, lysine‐specific demethylase 1.