Figure 4

Confirmation of ALK and MELK inhibition effects on ERα levels and cell proliferation in MCF-7 and MDA-MB-361 cell lines. Western blot analyses of ERα expression levels in MCF-7 and MDA-MB-361 cells treated with either MELK esiRNA (a,a’) or ALK esiRNA oligonucleotides for 24 h (c,c’), as well as with indicated doses of the MELK inhibitor MELK-8a (MELKin) (b,b’) or the ALK inhibitor AP26113 (AP) (d,d’) for 48 h. Representative blot images are shown. (a’’–d’’) Densitometric analyses of the corresponding blots. In panels (a’’) and (c’’), significant differences were calculated using the ANOVA test, and * indicates differences compared to control (CTR) samples (**p < 0.01, ****p < 0.0001), while ° indicates differences compared to esiRNA-treated samples (°°p < 0.01). In panels (b’’) and (d’’), significant differences were calculated for each dose in the different cell lines in the Student’s t-test test, and * represents a p-value < 0.05, *** represents p-values < 0.001, and **** represents p-values < 0.0001. (e) The inhibitor concentration 50 (IC₅₀) was calculated for both MCF-7 and MDA-MB-361 cells treated with different doses of the MELK inhibitor MELK8a (MELKin) for 7 days. Each dot represents an experimental replica. Significant differences were calculated using the Student’s t-test test, and **** indicates a p-value < 0.0001. (f) The inhibitor concentration 50 (IC₅₀) was calculated for both MCF-7 and MDA-MB-361 cells treated with different doses of the ALK inhibitor AP26113 (AP) for 7 days. Each dot represents an experimental replica. Significant differences were calculated using the Student’s t-test test, and * indicates a p-value < 0.05. Blots were cut prior to hybridization with antibodies during blotting. Images of all replicate blots are presented in Supplementary Fig. S2.