Fig. 5

SNHG17 interacted with the PGK1 protein in THP-1 cell-derived TAMs. A RNA-binding proteins potentially binding to SNHG17 tapped by RNA pull-down experiments. B Up: Immunoblotting to validate the interaction between PGK1 protein and biotinylated SNHG17. Down: PGK1 protein expression level in TAMs and NTRMs from PDAC patients. (n = 4) (C-D) PGK1 antibody was used for RIP-qPCR to determine whether SNHG17 binds to PGK1 in THP-1 cell-derived TAMs (co-cultured with PANC-1 cells (C) and PATU-8988 cells (D)). E Prediction of SNHG17 and PGK1 protein binding regions by catRAPID. F SNHG17 secondary structure analyzed by RNAfold web server. The fragments of SNHG17 are shown in red boxes. G Immunoblot analysis of the ability of PGK1 to directly bind to biotinylated SNHG17 truncations. H Schematic representation of the structural domain of the PGK1 protein. I Flag RIP-qPCR analysis revealing binding levels of various truncations of PGK1 to SNHG17 in THP-1 cells co-cultured with PANC-1 cells or PATU-8988 cells. (J) Representative images of the colocalization of SNHG17 and PGK1 in THP-1 cell-derived TAMs. K-L WB and qPCR analyses of SNHG17 and PGK1 in THP-1 cells in which PGK1 was knocked down following co-culture with PANC-1 cells (K) or PATU-8988 cells (L). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001