Research Paper Volume 15, Issue 9 pp 3230—3248

Figure 5. PTEN-PI3K-AKT axis and PD-L1 expression in BCC subsets. Western blots for PTEN and pPTEN (A), pAKT and AKT (B) and PI3K (C). Representative bands for the Western blots are shown with the mean±SD of normalized band densities for five biological replicates. (D) Gating strategy for BCC subsets were used for PD-L1 by flow cytometry. R5 represents Oct4^hi; R6, Oct4a^med; R7, Oct4a^lo. The voltage was consistently applied to isolate the various BCC subsets. (E) The dot blots represent three different independent experiments to assess the expression of Msi-1 and/or PD-L1 in BCCs shown in `D’. The isotype for these studies are shown below with arrows pointing to the experimental plots. (F) Western blot for PD-L1 with extracts from Msi-1 KD BCCs or vector control. The mean normalized band densities for three independent Western blots are shown at right (±SD). (G) Flow cytometry was conducted for PD-L1 with Msi 1 knockdown BCCs. The cells were gated as depicted in `D’. The grey region depicts fluorescence of isotype control. The histogram represents three independent studies with the percentages of each subset stated in the quadrants. * p<0.05 vs vector transfectants.