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. 2015 Nov 4;10(11):e0141988. doi: 10.1371/journal.pone.0141988

Fig 4. DMPB stimulates melanin synthesis by activating USF1.

Fig 4

(A) B16F10 cells were treated with 30 μM of DMPB for the indicated time periods. Cytoplasmic and nuclear fractions were isolated and analyzed by Western blotting. (B) B16F10 cells were transfected with si-CON or si-MITF for 48 hr. MITF protein levels were analyzed by Western blotting in the absence or presence of 1 μM α-MSH for 3 hr. (C) Tyrosinase protein levels were analyzed by Western blotting in the absence or presence of 30 μM DMPB for 48 hr (top panel) and the melanin content was measured by absorbance at 405 nm (bottom panel). The mean percentages of melanin content are shown. **, p < 0.01 versus DMSO treated cells. (D) B16F10 cells were treated with 30 μM of DMPB for the indicated time periods in the presence of 1 μM α-MSH. MITF and p53 protein levels were analyzed by Western blotting and MITF mRNA level was analyzed by RT-PCR. (E) B16F10 cells were treated with DMPB for 3 hr, and Immunofluorescence analysis was performed with anti-USF1 antibody. Scale bar = 20 μm (F) B16F10 cells were transfected with si-con or si-USF1 and treated with DMPB (30 μM) for 48 hr. Indicated protein levels were analyzed by Western blotting (top panel) and the melanin content was measured by absorbance at 405 nm (bottom panel).**, p<0.01; *, p < 0.05 versus DMSO treated cells.