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. 2019 May 16;10(6):378. doi: 10.1038/s41419-019-1598-x

Fig. 5. PTBP2 regulates the translation of RUNX2 in CRC cells.

Fig. 5

a Insertion of the RUNX2, VEGFa, and COX-2 5′UTR sequences (410 bp, 1038 bp, and 134 bp, respectively) into the empty dicistronic vector pR-F. b Renilla (RL) and firefly (FL) luciferase activity analyses following transfection of LoVo cells with different concentrations (0 ng, 12.5 ng, 25 ng, and 50 ng) of PTBP2-expressing plasmid. c Biotinylated RUNX2, VEGFa, and COX-2 5′UTR RNAs bound to streptavidin beads were incubated with cytoplasmic extracts from LoVo cells. After extensive washing, RNA-bound proteins were analyzed by western blot using an anti-PTBP2 antibody. d Schematic representation of the RUNX2 and VEGFa fragments. The polypyrimidine tract (PPT) is indicated as a black ellipse. e–f Biotinylated RUNX2 and VEGFa 5′UTR RNAs or fragment RNAs bound to streptavidin beads were incubated with cytoplasmic extracts from LoVo cells. After extensive washing, RNA-bound proteins were analyzed by western blot using an anti-PTBP2 antibody. g–j qPCR and western blot analysis for RUNX2, VEGFa, and COX-2 in LoVo and SW620 CRC cells transfected with PTBP2-expressing plasmid