Fig. 6

POU2F1 activates LINC01564 transcription and promotes GC proliferation and invasion. A The RNA levels of POU2F1 (left) and LINC01564 (right) were detected by qPCR after POU2F1 overexpression and knockdown. B The protein levels of POU2F1, E-cadherin, Vimentin, and β-actin were analyzed using Western blot. Group a, b, and c represent NC, POU2F1 OE, and POU2F1 KD, respectively. C ChIP assay was conducted in SGC-7901 and MKN-45 transfected with POU2F1 OE or NC. Cells were divided into IgG and ChIP groups. In ChIP group, POU2F1 was pulled down and the promoter sequence of LINC01564 was detected using qPCR. IgG and Poly-II were used as the negative and positive controls, respectively. D Biotinylated DNA pull-down assay was conducted in SGC-7901 and MKN-45 cells using WT or MT LINC01564 promoter probe. SDS-PAGE gel and Western analysis of POU2F1 were performed. SGC-7901 and MKN-45 cells transfected with NC, POU2F1 (OE), POU2F1 (KD), POU2F1 (OE) + LINC01564 (KD) or POU2F1 (KD) + LINC01564 (OE). Then cells were subjected to MTT (E), invasion (F) and colony formation assays (H). Significant differences are indicated by *p < 0.05, **p < 0.01 and ***p < 0.001