Skip to main content
. 2016 Sep;28(9):1225–1236. doi: 10.1016/j.cellsig.2016.05.023

Fig. 3.

Fig. 3

Ral-Arf6 crosstalk in Ras-transformed MiaPaCa2 and T24 cells. (A–C) Western blot detection and quantitation of active Arf6 pulled down by GST-GGA3 (GGA3) and total Arf6 in the respective whole cell lysate (WCL) from MiaPaCa2 cells grown with low serum(A) stable adherent (SA) and suspended (Susp), (B) suspended control (CON) and RalA knockdown (RalAi) and (C) suspended control (CON) and RalB knockdown (RalBi). (D–G) Western blot detection and quantitation of active Arf6 from T24 cells stable adherent (SA) and suspended for 120 min (Susp) (D) with low serum and (F) with serum. Similar Active Arf6 levels were determined in control (CON), RalA knockdown (RalAi) and RalB knockdown (RalBi) T24 cells suspended for 120 min (E) with low serum and (G) with serum. (H) Western blot detection and quantitation of active Arf6 and active RalA from WTMEFs stable adherent (SA) and suspended for 120 min (SUS) in the absence (CON) and presence of G12V H-Ras (H-Ras) with low serum. Calculated percentage active Arf6 levels and RalA levels were normalized to respective SA/CON. Graphs represent mean ± standard error from a minimum of three and maximum of five independent experiments as indicated in each graph. Statistical analysis of normalized data was done using the two tailed single sample t-test and their significance represented (* p value < 0.05, ** p value < 0.01 and *** p value < 0.001).