Fig. 6

ROCKII inhibition promotes the generation and maturation of glucose-responding cells. a Scheme of the directed differentiation protocol. b–d Intracellular FCM (b), Confocal imaging (c) and qRT-PCR analysis (d) of INS ^w/GFP HES3-carrying vectors to express shROCKI, shROCKII or scrambled shRNA in the presence or absence of 10 ng/ml dox treatment. Primary human islets and H1152-treated cells were used as controls in d. Error bar is SEM. e The percentage of c-peptide content and fold induction of ROCKI-KD and ROCKII-KD cells after 30 mM KCl treatment. Fold induction was calculated by dividing the percentage of c-peptide content in the KCl treated condition by that of the untreated condition. N = 8–12 independent biological replicates. f The percentage of c-peptide content and fold induction of ROCKI-KD and ROCKII-KD cells in the presence of 2 mM D-glucose (LG) and 20 mM d-glucose (HG). Fold induction was calculated by dividing the percentage of c-peptide content in the HG treated condition by that of the LG treated condition. Scale bar: 25 µm. Primary human islets and H1152-treated cells were used as controls in d. H1152-treated cells were used as controls in e and f. N = 8–12 independent biological replicates. p-values were calculated by unpaired two-tailed Student’s t-test. *p < 0.05, **p < 0.01. N6.1, NKX6.1. The bottom and top of the box represent the first and third quartiles, the band inside the box represents the median. The ends of the whiskers represent the minimum and maximum of all the data