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. 2023 Dec 4;43(5):363–377. doi: 10.1038/s41388-023-02897-y

Fig. 6. Chemical inhibition of PRMT5 has selective toxicity for neuroblastoma cells compared to normal myofibroblast cells.

Fig. 6

A Cell viability measured 72 h after SK-N-BE(2)-C, KELLY, SH-SY5Y, SK-N-FI, and MRC-5 cells were treated with increasing concentrations (0–50 µM) of the PRMT5 inhibitor, JNJ-64619178. B SK-N-BE(2)-C and KELLY cells were treated with IC50 concentrations of JNJ-64619178 for 10 days (SK-N-BE(2)-C) or 14 days (KELLY), followed by colony formation assays. Quantification of colony forming assays was based on colony numbers. Differences in colony formation were compared to the vehicle control. All results represent n = 3 independent biological replicates, mean ± SEM, where the P value was determined through a one-way ANOVA, with Dunnett multiple comparison testing. C Representative western blot of SNRPD3 protein arginine methylation (methylated SNRPD3), SNRPD3, and PRMT5 expression following treatment with IC50 concentration of JNJ-64619178 in SK-N-BE(2)-C and KELLY cells for 24 and 48 h, followed by densitometry analysis, where difference in protein expression was compared to vehicle control. D Cell viability was measured 72 h after vehicle or Dox treated SHEP.tet21n cells were treated with increasing concentrations (0–50 µM) of JNJ-64619178. E Representative western blot of SNRPD3 protein arginine methylation, SNRPD3, PRMT5, and MYCN expression following treatment of vehicle or Dox treated SHEP.tet21n cells with IC50 concentrations of JNJ-64619178 for 24 h, followed by densitometric analysis of protein expression compared to vehicle control. F Cell viability was measured 72 h after vehicle or Dox treated BE(2)-C.shSNRPD3 #2 cells were treated with increasing concentrations (0–50 µM) of JNJ-64619178. G Representative western blot of SNRPD3 protein arginine methylation, SNRPD3, and PRMT5 expression following treatment of vehicle or Dox treated BE2C.shSNRPD3 #2 cells with IC50 concentrations of JNJ-64619178 for 24 h, followed by densitometry analysis of protein expression compared to vehicle control. Two-way ANOVA statistical test was performed for each concentration compared back to no drug control (at 0 μM) (****p < 0.0001). All results represent n = 3 independent biological replicates, mean ± SEM, where the P value was determined through a two-way ANOVA, with Tukey’s multiple comparison tests, unless stated otherwise.