Journal: Cancer Biology & Therapy

Article Title: ARHGAP40 suppresses clear cell renal cell carcinoma progression and is associated with altered glutamine-related metabolism and redox balance

doi: 10.1080/15384047.2026.2746137

Figure Lengend Snippet: ARHGAP40 associates with RhoA and is accompanied by reduced RhoA activity, PI3K/AKT signaling, and altered glutamine-related/redox measures. (A) Flag-based co-immunoprecipitation showing association of ARHGAP40 with RhoA. (B) Endogenous immunoprecipitation of RhoA followed by immunoblotting for ARHGAP40. (C) RhoA activity assay using a conformation-specific anti-RhoA-GTP antibody, with representative GTP-RhoA and total RhoA blots and quantification of the GTP-RhoA/total RhoA ratio. (D) Representative western blots and densitometric analyzes of p -AKT/AKT and p -PI3K/PI3K. (E) Intracellular glutamate levels. (F) Intracellular glutamine levels. (G) GSH/GSSG ratios. (H) Representative ROS flow-cytometry histograms and mean fluorescence intensity. Quantitative analyzes used n = 3 independent biological replicates and are shown as mean ± SEM. Two-group comparisons were performed using two-tailed Student's t-test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST and incubated overnight at 4 °C with primary antibodies: β -actin (CST, 4970, 1:1000), ARHGAP40 (MyBioSource, MBS9215052, 1:1000), AKT (CST, 9272, 1:2000), phospho-AKT Ser473 (CST, 4060, 1:1000), PI3K p85 (CST, 4257, 1:1000), phospho-PI3K p85 Tyr458 (CST, 4228, 1:1000), RhoA (CST, 2117, 1:2000), E-cadherin (CST, 3195, 1:2000), N -cadherin (CST, 4061, 1:2000), and vimentin (CST, 5741, 1:2000).

Techniques: Activity Assay, Co-Immunoprecipitation Assay, Immunoprecipitation, Western Blot, Flow Cytometry, Fluorescence, Two Tailed Test