Figure 4. Caveolin-1 is required for K-RAS function and localization.
4A. Intracellular levels of phospho-caveolin-1 in HCT116 isogenic system
4B. Phosphorylation of caveoilin-1 is necessary for caveolin-1 interactions with K-RAS signaling molecules Grb2 and SOS1.
HCT116-Caveolin-1-anti-sense cells were transfected with WT or mutant (Y14F) caveolin-1 and lysed in RIPA buffer 48 hours after transfection. Lysates were immunoprecipitated with indicated proteins and immune complexes were resolved on SDS-PAGE gels and probed for indicated proteins.
4C. K-RAS protein localization in caveolin-1 depleted Caco-2 isogenic system.
CacoKras#26 were transfected with scrambled or caveolin-1 siRNA 24 hour after subculture and fractionated into lipid rafts 48 hours after transfection as described in 61. Caveolin-1, K-RAS and flotilling-1 protein levels were determined by Western blotting. Figure is representative of three independent experiments
4D. K-RAS localization in HCT116 mock and HCT116 CAV-1 AS isogenic system.
HCT116-Mock and Caveolin-1 anti-sense cells were fractionated into lipid rafts.
K-RAS and flotilling-1 protein levels were determined by Western blotting. Figure is representative of three independent experiments