Fig 1: KPNA4 was a direct target of miR‐548I. (a) Putative miR‐548I binding site in the 3′‐UTR of KPNA4 is shown. (b), (c) Dual‐luciferase reporter assay was performed to confirm the binding site of KPNA4 and miR‐548I. (d), (e) RIP assay was used to test the combination of KPNA4 and miR‐548I. (f) The mRNA expression of KPNA4 and corresponding normal tissues were detected by qRT‐PCR. (g), (h) The protein level of KPNA4 in NSCLC patients and cell were measured by western blot. (i) Western Blot was used to detect the expressions of KPNA4 in NSCLC cells. KPNA4, karyopherin subunit α 4; MiR, microRNA; RIP, RNA immunoprecipitation; mRNA, messenger RNA; qRT‐PCR, quantitative real‐time polymerase chain reaction; NSCLC, non–small cell lung cancer
Fig 2: miR‐548I repressed proliferation and migration and induced apoptosis of NSCLC cells by regulating KPNA4. (a) The protein expression of KPNA4 was detected by western blot in NSCLC cells. (b), (c) Cell proliferation viability was measured by CCK‐8 assay and EdU assay. (d), (e) The influence of miR‐548I and KPNA4 on cell migration and invasion of NSCLC cells measured by transwell assay. (f) Tube formation assay for the angiogenesis of transfected cells. (g) Cell apoptosis was determined by flow cytometry assay. (h), (i) The expression of Bax and Bcl‐2 was determined by western blot. MiR, microRNA; NSCLC, non–small cell lung cancer; KPNA4, karyopherin subunit α 4; CCK‐8, cell counting kit‐8; EdU, 5‐ethynyl‐2′‐deoxyuridine
Fig 3: Circ_0008003 significantly affected tumor growth in vivo. (a), (b) The tumor volume (a) and weight (b) in mice were determined. (c), (d) The relative levels of circ_0008003 and miR‐548I in tumor tissues were determined by qRT‐PCR. (e) The relative levels of KPNA4 protein expression in tumor tissues were determined by western blot. (f) Immunohistochemistry staining was used to determine the expression of KPNA4, Bcl‐2 and Bax in tissues from mice. qRT‐PCR, quantitative real‐time polymerase chain reaction; KPNA4, karyopherin subunit α 4
Fig 4: Circ_0008003 positively regulated KPNA4 by sponging miR‐548I in NSCLC cells. Western blotting was used to detect KPNA4 protein expression in NSCLC cells. KPNA4, karyopherin subunit α 4; NSCLC, non–small cell lung cancer
Supplier Page from Abcam for Anti-KPNA4 antibody