Supplementary Figure 6: OTUD3 negatively regulates Akt signaling in a PTEN-dependent manner.

a,b, Quantitative RT-PCR analysis of indicated gene mRNA level in MCF7 (PTEN wild-type) (a) or BT549 (PTEN-null) (b) cells stably expressing the shRNA-Control or shOTUD3#1. Data were analysed using two-tailed unpaired Student’s t-test. Data are shown as mean ± s.d. n = 3 independent experiments. *: P < 0.05,**: P < 0.01, NS: no significance. c, Quantitative RT-PCR analysis of VEGF gene mRNA level in MCF7 and BT549 cells stably expressing shRNA-Control or shOTUD3 #1. Data were analysed using two-tailed unpaired Student’s t-test. Data are shown as mean ± s.d. n = 3 independent experiments. d,e, OTUD3 shRNA-transduced MCF7 or BT549 cells were serum-starved and treated with EGF for various times. Quantification of the phosphorylation levels of Akt, p38 and Erk protein relative to total Akt, p38 and Erk protein levels for Fig. 3f and g. Data are shown as mean ± s.d. n = 3 independent experiments (two-way ANOVA test). Statistics source data can be found in Supplementary Table 4.