Journal: BMC Cancer

Article Title: Bidirectional alteration of Cav-1 expression is associated with mitogenic conversion of its function in gastric tumor progression

doi: 10.1186/s12885-017-3770-y

Figure Lengend Snippet: Opposite effects of low and high Cav-1 on inhibitory phosphorylation of RAF. a Cav-1 inhibition of EGFR stability. WT-Cav-1- or shCav-1-expressing cells were exposed to EGF (10 ng/ml) and EGFR level was determined using an immunoblot assay. b Effect of Cav-1 on Ras activity. GTP-Ras levels were measured using Ras activity assay. c , d Opposite effects of Cav-1 on EGF-induced MEK1/2 phosphorylation. e , f Effect of Cav-1 on EGF-induced RAF phosphorylation. Stimulatory phosphorylation of A-RAF (P-S299), B-RAF (P-S445) and C-RAF (P-S339) and inhibitory phosphorylation of C-RAF (P-S289/296/301) were detected using antibodies specific to phospho-RAF isoforms

Article Snippet: Western analyses were performed using antibodies specific for Cav-1 (sc-894, Santa Cruz, CA), Cav-2 (610,685, BD bioscience, CA), EGFR (#4267, Cell Signaling, Danvers, MA), RAF (#2330, Cell Signaling), MEK1/2 (#9911, Cell Signaling), ERK (#9101, Cell Signaling), AKT (#4060, Cell Signaling), JNK (#4668, Cell Signaling), and β-tubulin (T8328, Sigma).

Techniques: Phospho-proteomics, Inhibition, Expressing, Western Blot, Activity Assay

Journal: BMC Cancer

Article Title: Bidirectional alteration of Cav-1 expression is associated with mitogenic conversion of its function in gastric tumor progression

doi: 10.1186/s12885-017-3770-y

Figure Lengend Snippet: Reciprocal regulation of the RAF-ERK feedback loop by Cav-1. a V alidation of the RAF-ERK feedback regulation. Cells were treated with the MEK inhibitor UO126 and its effect on the inhibitory phosphorylation level of C-RAF was determined. b , c Effect of ERK depletion on Cav-1 regulation of RAF, MEK, and ERK. Cells were co-transfected with si-ERK2 and WT-Cav-1 or si-Cav-1. d , e Effect of ERK depletion on Cav-1 regulation of EGF-induced cell growth. WT-Cav-1- or shCav-1-expressing sublines were transfected with si-ERK2 and incubated with EGF (10 ng/ml). Data represent means of triplicate assays (Bars, SD). f Schematic representation of the opposite functions of Cav-1 in the regulation of the RAF-ERK feedback phosphorylation loop in gastric cancer cells

Article Snippet: Western analyses were performed using antibodies specific for Cav-1 (sc-894, Santa Cruz, CA), Cav-2 (610,685, BD bioscience, CA), EGFR (#4267, Cell Signaling, Danvers, MA), RAF (#2330, Cell Signaling), MEK1/2 (#9911, Cell Signaling), ERK (#9101, Cell Signaling), AKT (#4060, Cell Signaling), JNK (#4668, Cell Signaling), and β-tubulin (T8328, Sigma).

Techniques: Phospho-proteomics, Transfection, Expressing, Incubation