Journal: Biology Direct
Article Title: H3K27ac-activated EGFR-AS1 promotes cell growth in cervical cancer through ACTN4-mediated WNT pathway
doi: 10.1186/s13062-021-00315-5
Figure Lengend Snippet: EGFR-AS1 activates ACTN4-mediated WNT pathway in CC cells. a The knockdown efficacy of sh-ACTN4#1/2 on was tested by qRT-PCR in CC cells. b Expression levels of WNT pathway downstream genes in sh-ACTN4#1 transfected cells were detected by qRT-PCR in CC cells. c The effect of ACTN4 deficiency on the nuclear translocation of CTNNB1 (β-catenin) was detected by subcellular fractionation assay. d TOP/FOP flash assay was performed to measure the activity of WNT pathway upon ACTN4 knockdown. e , f Detection of mRNA and protein levels of WNT pathway downstream genes in CC cells transfected with sh-EGFR-AS1#1 was detected by qRT-PCR and western blot assays. g , h The nuclear translocation of CTNNB1 (β-catenin) was observed in sh-EGFR-AS1#1 group and sh-NC group. i Impact of EGFR-AS1 silencing on WNT pathway activity was detected. j – l Cell proliferation, apoptosis and invasion in cell transfected with sh-NC, sh-EGFR-AS1#1 and sh-EGFR-AS1#1 + LiCl were assessed by EdU, flow cytometry and transwell assays. **P < 0.01
Article Snippet: Besides, TOP/FOP-flash reporter vectors were available to assay the Wnt/β-catenin signaling activity as guided by manufacturer (Addgene, Cambridge, MA).
Techniques: Quantitative RT-PCR, Expressing, Transfection, Translocation Assay, Fractionation, Activity Assay, Western Blot, Flow Cytometry