Figure 4: G1 arrest by different mechanisms enhances the transdifferentiation.

(a–d) Representative images of MRC5 cells reprogrammed in full media by ANLmp in the presence of fetal bovine serum (FBS) without (a) or with the CDK2 inhibitor SU9516 (SU, 5 μM for 48 h) (b). The percentage of TH⁺ or Tuj1⁺ cells in all cells (DAPI⁺) (c) and the number of TH⁺, Tuj1⁺ or DAPI⁺ cells per frame under × 10 lenses (d) were analysed at day 10. *P<0.05, unpaired, two tailed Student’s t-tests versus the preceding bar; n=6 wells from 3 independent experiments. (e–h) Representative images of MRC5 cells reprogrammed in full media by ANLmp in the absence of FBS without (e) or with the CDK inhibitor SU9516 (SU, 5 μM for 48 h) (f). The percentage of TH⁺ or Tuj1⁺ cells in all cells (DAPI⁺) (g) and the number of TH⁺, Tuj1⁺ or DAPI⁺ cells per frame under × 10 lenses (h) were analysed at day 10. (i–l) Representative images of MRC5 cells reprogrammed in full media by ANLmp in the presence of FBS without (i) or with the mTOR inhibitor Torin 1 (0.1 μM for 48 h) (j). The percentage of TH⁺ or Tuj1⁺ cells in all cells (DAPI⁺) (k) and the number of TH⁺, Tuj1⁺ or DAPI⁺ cells per frame under × 10 lenses (l) were analysed at day 10. *P<0.05, unpaired, two tailed Student’s t-tests versus the preceding bar; n=6 wells from 3 independent experiments. Representative images of MRC5 cells reprogrammed in full media by ANLmp in the absence of FBS without (m) or with the mTOR inhibitor Torin 1 (0.1 μM for 48 h) (n). The percentage of TH⁺ or Tuj1⁺ cells in all cells (DAPI⁺) (o) and the number of TH⁺, Tuj1⁺ or DAPI⁺ cells per frame under × 10 lenses (p) were analysed at day 10. Scale bar, 100 μm.