Figure 10.

Cellular effects of compounds 12f–12h. (a, b) The viability of HEK293T cells was assessed by measuring the mitochondrial-dependent reduction of MTT to formazan, with respect to DMSO, after treatment with compounds 12f–12h at three different concentrations (10, 50, and 100 μM) for (a) 24 h and (b) 72 h. Data are reported as the mean ± SD of four independent experiments. (c, d) Western blot analyses were performed (a) on lysates from HEK293T cells after treatment with compounds 12f–12h at 10 and 50 μM for 24 h and (d) on lysates from MCF7 cells after treatment with compound 12h at 10 and 50 μM for 4 and 8 days. Methylation was detected by immunoblotting with a pan-PRMT4 substrate antibody (PRMT4^sub; see the main text).¹⁰² Total histone H3 (c) or actin (d) was used to check for equal loading. The cell-permeable PRMT4 inhibitor TP064 (10 μM) was used as a reference compound. (e, f) Relative proliferation of (e) HEK293T and (f) MCF7 cells with different concentration of 12h for different time points. The medium was changed at day 4. All the data points represent the relative viability normalized to day 0. The error bars represent the standard deviation of three biological replicates performed at each time point.