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. 2014 Jul 23;5(15):6387–6403. doi: 10.18632/oncotarget.2201

Figure 3. HMGB1 depletion was associated with p21 upregulation in a p53 independent manner.

Figure 3

(A) The mRNA levels of p21 from A375, G361 or SK-MEL-28 cell line expressing the indicated shRNA were determined by qRT-PCR. The numbers were mean ± SD from three experiments performed in triplicate. Bars, SD; *p<0.05. (B) The cells as in A were subjected to Western blot analysis using the indicated antibodies. (C) A375 and G361 cells were transfected with p53 siRNA (si-p53) or control siRNA (si-c), with or without stable HMGB1 depletion. The cells were analyzed via Western blot using the indicated antibodies. (D) H1299 (p53-null) cells were transfected with either control (sh-c) or HMGB1 (sh-2) shRNA and the cells were analyzed for the expression of HMGB1 and p21. (E) The expression of HMGB1 was knocked down in SK-MEL-28 cells and the protein (left panel) and mRNA levels (right panel) were analyzed. Representative results from one of three independent experiments performed in triplicate are shown. Bars, SD; *p<0.05. (F) Growth curve of SK-MEL-28 expressing shRNA HMGB1 (sh-1 or sh-2) or the control shRNA (sh-c) was determined. The numbers were mean ± SD from three experiments performed in triplicate. Bars, SD; *p<0.05. (G) Cells as in F were subjected to colony formation assay. Representative colony images are shown (left panel). The bar chart shows a quantitative analysis of the colony formation assay (right panel). The numbers were mean ± SD from three experiments performed in triplicate. Bars, SD; *p<0.05.