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. 2020 Jun 9;12(6):1500. doi: 10.3390/cancers12061500

Figure 6.

Figure 6

sPAH inhibits doxorubicin, vemurafenib and cholesterol efflux by directly binding to Ptch1. (A) Direct binding of sPAH to Ptch1. Membranes from yeast expressing hPtch1 or hSmoothened proteins labeled with tris-NTA-NT647 fluorescent probe were incubated with sPAH and analyzed by microscale thermophoresis (MST). Kd reported is the mean of the Kd values determined in 3 independent experiments. (B) sPAH inhibits BODIPY-cholesterol efflux. A375 cells were seeded on coverslips and incubated with BODIPY-cholesterol (Bo-chol) for 2 h. Three coverslips were fixed for Bo-chol loading control, while the others were incubated with DMSO or sPAH and fixed. (C) Vemurafenib inhibits doxorubicin efflux. A375 and WM9R cells were seeded on coverslips, incubated with dxr in the absence or the presence of vemurafenib for 1 h and fixed. BODIPY and dxr fluorescence were imaged and quantified using ImageJ software for about 100 cells per condition per experiment. All data presented are the mean ± SEM of 3 independent experiments. Significance is attained at p < 0.05 (*) (**: p < 0.005).