Fig. 5

RP11-296E3.2 interacts with YBX1 to promote CRC progression. A Model of the RNA pulldown experimental workflow (left panel). The specific association between RP11-296E3.2 and YBX1 in HCT116 cells was validated using an RNA pulldown assay. WB was performed to verify the RNA pulldown assay results. The RP11-296E3.2 antisense sequence was used as a control (right panel). B RNAalifold predicted that RP11-296E3.2 contains 5 stable stem‒loop structures. The inset (framed in red) shows the YBX1 binding stem‒loop structures in RP11-296E3.2. C RNA pulldown followed by WB analysis was performed with a series of deletion constructs of RP11-296E3.2. D Model of the RIP assay workflow (left panel). RIP assays were performed with YBX1. qRT‒PCR analysis revealed that RP11-296E3.2 was significantly enriched in the anti-YBX1 immunoprecipitates relative to the IgG control immunoprecipitates (right panel). E RIP assays were performed to quantify the expression of RP11-296E3.2 in cells transfected with YBX1 and its deletion mutants. The data are presented as the means ± SDs. Statistical significance was assessed using one-way ANOVA followed by Tukey’s test for multiple comparisons. ****P < 0.0001