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. 2015 Feb 12;6(2):e1637. doi: 10.1038/cddis.2015.2

Figure 2.

Figure 2

Elevation of PAK1 kinase activity displays stem-like characteristics in RCC cells. (a) Western blot analysis of PAK1, p-PAK1 (T423) and GAPDH for HKC, OS-RC-2, 786-O and ACHN cells. GAPDH was used as a loading control. The numbers shown are the ratios of the intensities of the bands for p-PAK1, divided by the intensities of the bands for PAK1, normalized to 1.00 for HKC. (b) Western blot analysis of PAK1 and p-PAK1 (T423), RAF1, p-RAF1(S338), MEK1, p-MEK1(S298) and GAPDH for HKC, 786-O and OS-RC-2 cells stably transfected with PAK1 T423E. GAPDH was used as a loading control. The numbers shown are the ratios of the intensities of the bands for pPAK1(T423), p-RAF1(S338) and p-MEK1(S298), divided by the intensities of the bands for GAPDH, RAF1 and MEK1, respectively, normalized to 1.00 for cell lines without PAK1 T423E transfected. (c) Analysis of self-renewal of HKC, 786-O and OS-RC-2 cells stably transfected with control (empty vector) or PAK1-T423E. Data are represented as means±S.D. of triplicate experiments. *P<0.05 versus control. (d) Flow cytometry analysis of ALDH1, CD73 and CD146 for HKC, 786-O and OS-RC-2 cells stably transfected with control or PAK1 T423E from generations G1 to G3 spheres. Data are represented as means±S.D. of triplicate experiments. *P<0.05 versus control