Fig. 4

PANC-1 cell stemness is regulated by miR-384 thought targeting ACVR1. a, miR-384 expression and mRNA expression of ACVR1 and CSC markers (Oct4, ABCG2, Nestin, CK19 and CD133) as determined by RT-qPCR; b, grey value of Oct4, ABCG2, Nestin, CK19, CD133, ACVR1 and GAPDH protein bands in as measured by western blot analysis; c, protein expression of Oct4, ABCG2, Nestin, CK19, CD133 and ACVR1; d, micrograph of sphere formation of PANC-1 cells (× 200); e & f, statistical chart of sphere diameter and spheres per 100 cells of PANC-1 cells as determined sphere formation assay; g, statistical chart of monoclonal formation rate as evaluated by clone formation assay; h, proliferation of PANC-1 cells as assessed by EdU staining (× 200); i, statistical chart of cell proliferation; j, micrograph of the number of invasive cells as determined by Transwell assay (× 200); k, statistical chart of cell invasion; l, the mRNA expression of ACVR1 in PANC-1 cells as determined by RT-qPCR; m, micrograph of sphere formation of PANC-1 cells (× 200); n & o, sphere diameter and spheres per 100 cells of PANC-1 cells as determined by sphere formation assay; p, monoclonal formation rate as evaluated by clone formation assay; ^*, p < 0.05 vs. the miR-384 mimic-NC and miR-384 inhibitor-NC groups; ^#, p < 0.05 vs. the miR-384 inhibitor group. All the above data were measurement data and expressed as mean ± standard derivation. The t-test was performed for comparison between two groups. The experiment was repeated three times. PC, pancreatic cancer; miR-384, microRNA-384; NC, negative control; RT-qPCR, reverse transcription quantitative polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ACVR1, activin receptor A type I; ABCG2, ATP-binding cassette subfamily G member 2