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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: J Immunol. 2014 Jun 30;193(3):1162–1170. doi: 10.4049/jimmunol.1303162

Figure 6. mTORC2 disruption diminished CD147 expression on the surface of mouse embryonic fibroblasts or immortalized T-cells.

Figure 6

A. Wild-type (+/+) and SIN1-deficient (−/−) MEFs were harvested, stained for CD147 and propidium iodide (PI) followed by flow cytometric analysis of live cells (PI−). MFI of CD147 was plotted from 4 distinct experiments. B. Lysates of wild-type and SIN1-deficient MEFs were incubated for 1 hr in presence or absence of glycosidases followed by CD147 immunoblotting. High (HG) or low (LG) glycosylated or non-N-glycosylated (Non #) CD147 forms are indicated. Shown is a representative experiment out of three C. Lysates from MEFs were subjected to pull down assays with the indicated lectin agaroses. The means and SEM of fold changes of CD147 HG as compared to the expression in wild-type cells (lane 1) were indicated below each blot. Shown is a representative experiment out of three. D-E. Jurkat T-cells were transfected with rictor (kd) or scrambled (+/+) shRNAs. Jurkat cells, peripheral T-cells and thymocytes were (D) lysed and the phosphorylation and total protein expression were analyzed by immunoblotting. The mean and SEM of fold changes as compared to protein expressions in wild-type Jurkat cells (lane 2) were indicated below each blot. Shown is a representative experiment out of four, or (E) Cells were stained for TCRβ, and CD147 expression followed by flow cytometric analysis (n= 4).