Fig 1: ARPP19 is the downstream gene of miR-26b-5p in CRC cells. (a) The microT, miRanda, miRmap, PITA, and TargetScan databases jointly predicted underlying target genes of miR-26b-5p. (b) The mRNAs downregulated upon HCG11 knockdown and miR-26b-5p overexpression were screened as the candidates. (c) Expressions of 5 candidate mRNAs in CRC cell lines relative to NCM-460 cells were measured via qRT-PCR. (d) Western blot analyzed the expression of ARPP19 in different cell lines. (e) The binding sites between miR-26b-5p and ARPP19 were acquired after ENCORI prediction. (f) Luciferase reporter assay was done to verify the interaction between miR-26b-5p and ARPP19. (g) The existence of HCG11, miR-26b-5p, and ARPP19 in anti-AGO2 was detected by RIP assay. (h) ARPP19 expression was assessed via qRT-PCR in cells with transfection of sh-HCG11#1 or sh-HCG11#1+miR-26b-5p inhibitor. ∗∗p < 0.01.
Fig 2: HCG11 exacerbates CRC cell malignant behaviors through modulating the miR-26b-5p/ARPP19 axis. (a) The overexpression efficiency of pcDNA3.1-ARPP19 was assessed through qRT-PCR. HCT15 and HT-29 cells transfected with sh-NC, sh-HCG11#1, sh-HCG11#1+miR-26b-5p, or sh-HCG11#1+pcDNA3.1-ARPP19 were used for functional assays in a rescue manner. (b, c) CRC cell proliferation was evaluated by EdU assay and colony formation assay. (d, e) The cell apoptosis ability was measured via TUNEL assay and JC-1 assay. (f, g) The cell migration and invasion abilities were detected via transwell assay. ∗∗p < 0.01.
Fig 3: ARPP19 knockdown inhibits CRC cell malignant behaviors. (a) qRT-PCR measured the changes in ARPP19 expression after CRC cells were transfected with sh-NC, sh-ARPP19#1, or sh-ARPP19#2. (b, c) CRC cell proliferation was assessed via EdU assay and colony formation assay. (d, e) The apoptosis capability of transfected cells was evaluated by TUNEL and JC-1 assays. (f, g) Cell migration and invasion abilities were assessed via transwell assay. ∗∗p < 0.01.
Supplier Page from Abcam for Anti-ARPP-19 antibody