Fig. 3.

CBSLR physically interacts with YTHDF2 and modulates CBS level. (A) Schematic overview of in vitro RNA Antisense Purification and used for identification of proteins interacting with CBSLR. (B) RNA pull-down assay was performed to confirm the association between YTHDF2 and CBSLR. (C) RIP experiments shows the binding of CBSLR with YTHDF2. (D) RNA pull-down using sequentially deleted CBSLR fragments demonstrates the binding segment of CBSLR with YTHDF2. (E) shRNA-NC or shRNA-CBSLR MKN45 or MKN28 cells were cultured under normoxic or hypoxic conditions for 24 h. The mRNA and protein levels of CBS were determined using qRT-PCR or western blot analysis. (F) MKN45, and MKN28 cells stably infected with lentiviruses encoding lncRNA-CBSLR or NC were cultured under normoxic or hypoxic conditions for 24 h. The mRNA and protein levels of CBS were determined using qRT-PCR or western blot analysis. (G) The shRNA-NC and shRNA-CBSLR-1-MKN45 cells were cultured under normal or hypoxic conditions (24 h). m6A RIP-qPCR analysis of CBS or NANOG (positive control) mRNA was performed. (H) Schematic illustrations of m6A motifs positions of CBS mRNA.