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. 2015 Aug 26;6:8093. doi: 10.1038/ncomms9093

Figure 3. CAV1 regulates the transcriptional activity of β-catenin.

Figure 3

(a) TOP-FLASH activity in Hs944T cells in the presence of GFP, PTEN and/or CAV1 β-catenin (BCAT). (b) Similarly, TOP-FLASH activity was measured in the same cells post transfection of siRNA directed against negative control (siSCR), CAV1 (siCAV1), β-catenin (siBCAT). (c) Activity of p16INK4A::luciferase reporter was evaluated post transfection in Hs944T human melanoma cells with GFP, PTEN and/or CAV1 BCAT, and (d) with siSCR, siCAV1 and siBCAT. (a–d) All p16INK4A::luciferase and TOP-FLASH reporter assays were evaluated in the presence of an internal control (Renilla luciferase). (e,f) p16 mRNA level as measured by quantitative reverse transcriptase–PCR (fold change), following overexpression of GFP, PTEN and/or CAV1 BCAT, or knockdown of CAV1 and BCAT, with appropriate controls. (g) Eighteen human melanoma tumours were stained for CAV1, PTEN and p16. PTEN and p16 were absent and CAV1 was present for patient 1. Opposite observation was performed for patient 2. Stromal and endothelial cells were used as positive control for PTEN and CAV1, respectively. Scale bar, 100 μm for all panels. Error bars represent s.d. *P-value <0.05, **P-value <0.01 and ***P-value <0.001. Statistical significance was determined by Mann–Whitney test. Each experiment was performed in eight and three biological triplicates for a–d, e and f respectively.