Figure 3.

CASC21 bound to POU5F1B to synergistically up-regulate HGH1

a.CASC21 was co-expressed with POU5F1B based on GEPIA database.b-c.POU5F1B expression in CRC tissues versus adjacent normal tissue and in CRC cell lines versus normal FHC cells was evaluated through RT-qPCR.d.A high possibility of the interaction between CASC21 and POU5F1B protein was predicted via RPISeq based on the Random Forest (RF) and Support Vector Machine (SVM) classifiers. e.RNA pull down and western blot was conducted to verify the binding ability between POU5F1B and CASC21, antisense CASC21 and bio-PCAT2 serving as negative control. f.RIP assay was carried out to examine CASC21 enrichment in anti-IgG and anti-POU5F1B groups. g. RT-qPCR and western blot were conducted to detect the influence of CASC21 depletion on POU5F1B expression. h. RT-qPCR was used to detect the influence of POU5F1B depletion on CASC21 expression. i. Six genes were predicted to be co-expressed with both CASC21 and POU5F1B. j-k. Influence of respective or combined knockdown of CASC21 and POU5F1B depletion on the expression and protein levels of HGH1 was tested by RT-qPCR and western blot. *P < 0.05, **P < 0.01. The symbol ‘n.s’. means no significance.