Extended Data Fig. 2: RAB22A-NeoF1 is dominant in ZOS and ZOS-M cells.
From: Chromosomal translocation-derived aberrant Rab22a drives metastasis of osteosarcoma
a, The protein levels of each RAB22A-NeoF1-6 in U2OS cells transiently transfected with the indicated plasmids with or without 200 nm Bafilomycin A1 and 10 μm MG132 for 6 h. Vector, vector-only control. b, The reads of RAB22A-NeoF1-6 in both ZOS and ZOS-M cell lines. c, The detection of endogenous RAB22A-NeoF2/3/4/5/6 proteins by Western blotting in the indicated stable cell lines, as described in methods (Note: The Western blotting using anti-RAB22A-N followed by IP with anti-RAB22A-N was performed with the cut membranes below the molecular weight of 15 KDa to avoid the effects of wild type RAB22A). Data in a,c are representatives of n =3 biologically independent experiments. sgNC, negative control sgRNA. d, The amino acid sequence of RAB22A-NeoF1 full-length protein. Residues corresponding to the inverse DOK5 sequences are shown in red. e, Quantification analysis of mRNA levels of RAB22A-NeoF1 in the indicated osteosarcoma cell lines by real-time PCR. Mean ± s.d. of n = 3 biologically independent experiments. p values are shown. Two-tailed Student t-test. f, FISH analyses in ZOS and ZOS-M cells, as described in the methods. Red: RAB22A, Green: DOK5, yellow: the fusion of RAB22A and DOK5 indicated by arrows. Data are representatives of n = 3 biologically independent experiments. Statistical source data are provided in Source Data Extended Data Fig. 2.
