(A) Left: RT-PCR and Western blot results show increased HMGCL expression in human melanoma cells expressing BRAF V600E/D compared to cell expressing BRAF WT. Right: Western blot results of HMGCL expression in Mel-ST cells with FLAG-BRAF WT or V600E. Data are mean±s.d. n=3 each; p values were obtained by a two-tailed Student's t test.
(B) HMGCL immunohistochemistry. Left: Positive staining of HMGCL was determined by histochemical score (H score = 3 × percentage of strong staining + 2 × percentage of moderate staining + 1 × % of weak staining + 0 × % of no staining; score range 0~300). Representative IHC staining images for 0 (WT; no staining), 1+ (WT; weak staining), 2+ (V600E; moderate staining), and 3+ (V600E; strong staining) scores of human melanoma tissue samples are shown (20×). Right: H scores are presented by Box and Whisker plots. Medians, interquartile, maximum, and minimum are shown.
(C-E) Western blot and RT-PCR results show increased HMGCL expression with increased MEK1 and ERK phosphorylation in human primary melanoma (C) and hairy cell leukemia tissue samples (D-E). PB: peripheral blood; BM: bone marrow. Data are mean±s.d.; n=3 each; p values were obtained by a two-tailed Student's t test.
Also see Figure S2.