Fig. 4
From: Circ-HuR suppresses HuR expression and gastric cancer progression by inhibiting CNBP transactivation
CNBP promotes HuR expression, growth, and aggressiveness of gastric cancer. a Schematic illustration of dCas9-based CRISPRi for CNBP and small guide RNA (sgRNA) targeting region. b Dual-luciferase assay revealing the promoter activity of HuR in AGS and MKN-45 cells stably transfected with empty vector (mock), CNBP, CRISPRi-Scb, CRISPRi-CNBP #1, or CRISPRi-CNBP #2. (c) and (d) Real-time qRT-PCR (c, normalized to β-actin, n = 5) and western blot (d) assays showing the transcript and protein levels of HuR in AGS and MKN-45 cells stably transfected with mock, CNBP, CRISPRi-Scb, CRISPRi-CNBP #1, or CRISPRi-CNBP #2. e MTT colorimetric assay indicating the viability of AGS and MKN-45 cells stably transfected with mock, CNBP, CRISPRi-Scb, or CRISPRi-CNBP #1. f and g Soft agar (f) and matrigel invasion (g) assays showing the in vitro growth and invasion of AGS and MKN-45 cells stably transfected with mock, CNBP, CRISPRi-Scb, or CRISPRi-CNBP #1. ANOVA analyzed the difference in (b, c and e-g). *P < 0.01 vs. mock or CRISPRi-Scb. Data are shown as mean ± SEM (error bars) and representative of three independent experiments in (b-g)