Figure 6. Biochemical analysis of mTORC1- and mTORC2-dependent signaling in response to UVB. iRicKO cells were treated with vehicle or with 4OHT for 3 days to induce rictor recombination. On day four, cells were exposed to 20 mJ/cm<sup>2</sup> UVB, and harvested in 1× sodium dodecyl sulfate sample buffer at the indicated time points for immunoblot analysis. For each protein analyzed, samples from both treatment groups are loaded onto the same gel to allow direct comparison of protein expression levels. (A) Expression of mTORC1 signaling intermediates over a time course from T = 0 to T = 24h after UVB exposure. (B) Expression of mTORC2 signaling intermediates over the same time course. Quantitation of blots is shown in Supplementary Figures 1A and B is available at Carcinogenesis Online. Phosphorylated proteins were normalized to their corresponding total protein on the same gel and other proteins were normalized to either Lamin B1 or Tubulin on the same gel, as described in the legend for Supplementary Figure 2, available at Carcinogenesis Online. One Lamin B1 blot and one Tubulin blot are shown for reference. Results are representative of 2–3 independent experiments. From: Conditional disruption of rictor demonstrates a direct requirement for mTORC2 in skin tumor development and continued growth of established tumors Carcinogenesis. 2015;36(4):487-497. doi:10.1093/carcin/bgv012 Carcinogenesis | © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com 1