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. 2019 Feb 21;116(10):4548–4557. doi: 10.1073/pnas.1813802116

Fig. 1.

Fig. 1.

Down-regulation of FZD3 inhibits melanoma cell proliferation and colony-forming capacity. (A) Expression of FZD3 was analyzed by qRT-PCR following transduction of melanoma M727, M1626, and M525 cells with the control shRNA (shSC) or FZD3-specific shRNA (shFZD3-2 and shFZD3-3). Error bars represent SEM of duplicate experiments with three replicates each. (B and C) Immunofluorescence analysis of FZD3 protein expression in the indicated melanoma cells transduced with control shRNA (shSC) or shFZD3. The y axis indicates relative FZD3 protein fluorescence intensity. Red color indicates positive FZD3 staining. (Scale bars, 50 μm.) (D) Proliferation kinetics of shFZD3 and shSC-transduced melanoma cells, M727, M525, and M1626 quantified using Resazurin assay. Fluorescence values were normalized to the background readings collected from the wells that had no growing cells but contained growth culture media and Resazurin reagent. (E) Numbers of colonies formed by M727, M525, and M1626 melanoma cells following their transduction with the control shRNA (shSC) or shFZD3. Error bars represent SEM of duplicate experiments with two replicates each. *P < 0.05, **P < 0.005, ***P < 0.0005. (F) Representative images of Crystal violet-stained plates demonstrating colony growth potential of indicated melanoma cell lines transduced with the control shRNA (shSC) or shFZD3.