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. 2012 Aug 27;209(9):1689–1702. doi: 10.1084/jem.20101355

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Figure 4. — The EGFR autocrine loop is intact in RAS-transformed MyD88-deficient keratinocytes. (A) Tritiated thymidine incorporation was measured in control and v-ras^Ha–transduced WT keratinocyte cultures treated with IL-1ra for 3 d. Data shown are representative of three independent experiments, and bars represent the mean ± SEM value of four replicates. *, P < 0.05 between v-ras^Ha and control; #, no significant difference between v-ras^Ha − IL-1ra and v-ras^Ha + IL-1ra. (B) Total cell extract from primary keratinocytes transduced for 3 d with v-ras^Ha in the presence or absence of IL-1ra (Anakinra) was analyzed by Western blotting for phospho-EGFR (Tyr1068), total EGFR, and actin as loading control. The picture is representative of three independent experiments. (C) Real-time PCR analysis of Btc (betacellulin), Areg (amphiregulin), and Hbegf (heparin-binding EGF-like growth factor) in control and v-ras^Ha–transduced keratinocytes (WT and MyD88^−/−) 3 d after RAS transduction. (D) Real-time PCR analysis of Hras1 (H-ras) in control and v-ras^Ha–transduced WT and MyD88^−/− keratinocytes. (C and D) Data shown are representative of three independent experiments, and bars represent the mean ± SEM of three replicates. (E) Total cell extract from MyD88^−/− and IL-1R^−/− primary keratinocytes and their respective controls transduced for 3 d with v-ras^Ha were analyzed by Western blotting for phospho-ERK, total ERK, H-ras, and actin as loading control. The picture is representative of two independent experiments. (B and E) Molecular mass is indicated in kilodaltons.