Extended Data Fig. 4: RAB22A-NeoF1 specifically activates RhoA, and depletion of RhoA abolishes the functions of RAB22A-NeoF1 in vitro.
From: Chromosomal translocation-derived aberrant Rab22a drives metastasis of osteosarcoma
a, The partial list of proteins from the mass spectrum using cells stably expressing RAB22A-NeoF1-SFB. The unique and total peptide numbers for the indicated proteins are shown. b, c, e, 293T cells co-transfected with the indicated plasmids for 48 h were subjected to the indicated assays. d, The detection of Rho-GTP and membrane RhoA (RhoA-MEM) in the indicated stable cell lines by Western blotting. Vector, vector-only control. f, The co-IPs were performed using ZOS cells with IgG, mAb RAD5-8 or anti-RhoA at their endogenous levels, and were analyzed by Western blotting using hRAD5-8-V1-R5. g, The indicated proteins were analyzed by Western blotting in the indicated stable cells. Data in b-g are representatives of n = 3 biologically independent experiments. h, i, Quantification analyses of the migration and invasion (h) or adhesion (i) assays using the indicated stable cell lines. Vector, vector-only control. j, k, Quantification analyses of the migration and invasion (j) or adhesion (k) assays using the indicated stable cell lines with or without the RhoA inhibitor CT04. Data in h-k are Mean ± s.d. of n = 3 biologically independent experiments. p values are shown. A two-tailed Student’s t-test was performed. Vector, vector-only control. Statistical source data are provided in Source Data Extended Data Fig. 4.
