Fig. 5

K55 sumoylation is essential for the transcriptional activity of SOX10 toward FOXD3. a Alignment of sumoylation motifs in SOX10 from different species and SOX9. Putative sumoylation sites were underlined and consensus sumoylation motif is shown. b HEK293T cells were co-transfected with plasmids expressing Flag-SUMO1 and one of the HA-SOX10 variants including WT, K55R, K357R, and 2KR. After 48 h, immunoprecipitation was performed with HA-tag antibody. Inputs and immunoprecipitates were analyzed by western blot. Relative levels of SOX10 sumoylation were quantitated by normalizing the intensities of the sumoylated bands against the non-sumoylated bands. c 1205Lu-TR cells transduced with lentiviruses carrying siRNA-resistant HA-SOX10 cDNA variants including K55R, K357R, and 2KR 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. d Same as (b) except that A375-TR cells were used. Uncropped images are shown in Supplementary Fig. 13