Fig. 4

Phosphorylation at T240 and/or T244 inhibits transcriptional activity of SOX10 toward FOXD3 and other targets. a 1205Lu-TR cells transduced with lentiviruses carrying siRNA-resistant, HA-tagged SOX10 cDNA variants including T240E, T244E, and EE were transfected with SOX10 siRNAs for 72 h in the absence or presence of 100 ng ml⁻¹ doxycycline. Cells were then lysed and analyzed by western blot on indicated proteins. Actin was used as a loading control. Quantitation of FOXD3 expression is shown below the corresponding blots. b Same as (a) except that A375-TR cells were used. c 1205Lu-TR (top) or A375-TR (bottom) cells transduced with either WT or EE HA-SOX10 lentiviruses were transfected with SOX10 siRNAs for 72 h with or without 100 ng ml^–1 doxycycline. Cells were then treated with 2 μM Vemurafenib for 24 h and lysed for total RNA isolation and qRT-PCR analysis. Average results from three independent experiments are shown. Error bars represent standard deviation. Significance was determined by ANOVA one-way test,*p < 0.05; **p < 0.01; ***p < 0.001. Uncropped images are shown in Supplementary Fig. 12