Fig. 7
From: PAN-cancer analysis of S-phase enriched lncRNAs identifies oncogenic drivers and biomarkers
SCAT7 interacts with hnRNPK and YBX1 to regulate cell proliferation and cell cycle progression. a Venn diagram showing SCAT7 interacting proteins in HeLa cells identified using ChOP-MS in two independent biological replicates. b RIP using hnRNPK or YBX1 antibody followed by qPCR for SCAT7. c Validation of SCAT7 interaction with hnRNPK and YBX1 by ChOP followed by immunoblotting in HeLa cells. LacZ and SCAT7 reverse biotinylated probes were used as negative controls. d MTT in HeLa cells after transfection with two siRNAs targeting hnRNPK or YBX1. e Cell cycle analysis upon hnRNPK and YBX1 KD in HeLa cells. f FGFR2 expression by real-time qPCR in hnRNPK and YBX1 KD HeLa cells. g Western blot of FGFR2, FGFR3, AKT, Ser 473 Phospho-AKT (p-AKT S473), ERK1/2, and Phoshpo-ERK1/2 (p-ERK 1/2) in hnRNPK and YBX1 KD HeLa cells. h, i ChOP followed by qPCR for SCAT7 enrichment at FGFR2 (h) and FGFR3 (i) promoters in HeLa cells. Four primer pairs were used to assess the occupancy at every 250 bp upstream of FGFR2 and FGFR3 TSS. j, k ChIP using hnRNPK or YBX1 antibody followed by qPCR for hnRNPK and YBX1 occupancy at FGFR2 (j) and FGFR3 (k) promoters in control and SCAT7 KD HeLa cells. l Interaction of SCAT7 with hnRNPK and YBX1 by SCAT7 ChOP followed by immunoblotting in A549 cells. m ChOP followed by qPCR for SCAT7 enrichment at FGFR3 promoter in A549 cells. n hnRNPK or YBX1 ChIP followed by qPCR depicting the occupancy of hnRNPK and YBX1 at the FGFR3 promoter in control and SCAT7 KD A549 cells. o SCAT7 ChOP followed by immunoblotting with hnRNPK or YBX1 antibody in Caki-2 cells. p ChOP followed by qPCR quantification of SCAT7 enrichment at FGFR2 promoter in Caki-2 cells. q hnRNPK ChIP followed by qPCR depicting the occupancy of hnRNPK at FGFR2 promoter in control and SCAT7 KD Caki-2 cells. Note that the data presented in b, d–f, h–k, m, n, p, and q represent mean values of three independent experiments and the statistical significance was derived using a two-tailed unpaired Student’s t-test. Data are plotted as mean ± SD (*p ≤ 0.05; **p = 0.01 ̶ 0.001; ***p < 0.001)
