Fig. 3

FAM167A binds to its receptor DSG1. A Surface FAM167A receptor (FAM167AR) staining of K562S and K562R cells using Myc-tagged FAM167A and an Alexa Fluor 488-conjugated anti-Myc antibody. B Silver staining of immunoprecipitated samples prepared from K562R cells using Myc-tagged FAM167A and an anti-Myc antibody bound to Protein G Sepharose® beads. C MS/MS spectrum and peptide sequence of the indicated band from B. D Co-immunoprecipitation analysis between FAM167A and DSG1 in K562R cells. E qRT-PCR analysis of DSG1 mRNA levels in K562S and K562R cells, and in K562R cells treated with 1 μM imatinib for 24 h. F Immunoblot analysis of DSG1 expression in K562S and K562R cells. G Surface staining of K562S and K562R cells using an anti-DSG1 antibody. H Immunofluorescence microscopy analysis of DSG1 expression in K562S and K562R cells. Nuclei were visualized using Hoechst 33342. Scale bars, 10 μm. GAPDH was used as an internal standard (F). IP, immunoprecipitation. MFI, mean fluorescence intensity. Data are representative of three (A, B, D-H) independent experiments (error bars, s.d. of duplicate (E) or triplicate (A, G) samples). Unpaired two-tailed t-test; **P < 0.01, ***P < 0.001. NS, not significant