Fig. 2

The interaction between YBX1 and G3BP1 in RCC. (a) Left panels: YBX1 and G3BP1 mRNA levels were quantified by real-time PCR with YBX1 knockdown in ACHN cells. Right panels: the expression of YBX1 and G3BP1 in YBX1-depleted ACHN cells were detected by western blot. (b) G3BP1 did not affect YBX1 mRNA and protein level in ACHN cells knockdown G3BP1. (c) The interaction network of G3BP1 protein with other proteins was identified using STRING database. Abbreviations: STRING, Search Tool for the Retrieval of Interacting Genes. (d) ACHN cell lysates were mixed with an anti-YBX1 antibody or IgG antibody that does not recognize YBX1. YBX1 and control immunoprecipitates were analyzed by western blot with anti-G3BP1 and anti-YBX1 antibodies. IP, immunoprecipitation. (e) Immunofluorescence assays in ACHN cells. The localizations of YBX1 and G3BP1 were detected by confocal laser scanning microscopy as indicated. Scale bar = 20 μm. (f) Left panels: schematic diagram of YBX1 domain. Right panels: Co-IP assay showing interaction of G3BP1 with YBX1 (aa 130–205). Statistically significant differences were indicated: *, P < 0.05. NS: no significant difference