Figure 2

MYC Box 0 and 2 domains are crucial for colony formation during reprogramming. (A) Schematic representation of WT c-MYC and MYCL protein. Black boxes show important domains for MYC function, including MB0, 1, 2, 3a (c-MYC only), 3b, 4, and basic-helix-loop-helix leucine zipper motif (bHLHLZ). The percentage of common amino acids in each MYC box domain between MYCL and c-MYC is shown (Identity%). The numbers on the right indicate amino acid lengths. (B) Number of iPSC-like and non-iPSC-like colonies transduced with EpiP including c-MYC-WT/mutants (left) or MYCL-WT/mutants (right) on day 21. Mean ± SD values are shown. n = 3, **p < 0.01, ***p < 0.001 and ****p < 0.0001 by ordinary one-way ANOVA and Dunnett’s test vs. WT. (C) Expression of TRA-1-60 ( +) HDFs and CD13 ( +) HDFs transduced with EpiP including c-MYC-WT/mutants (left) or MYCL-WT/mutants (right) on day 16. Mean ± SD values are shown. n = 3, *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 by ordinary one-way ANOVA and Dunnett’s test vs. WT. (D) Representative flow cytometry images for TRA-1-60 and CD13 for HDFs transduced with EpiP including c-MYC-WT/ΔMB0 or MYCL-WT/ΔMB0 10 and 21 days after the transduction. Numbers indicate the expression percentage of each quadrant. (E) Proliferation of HDFs transduced with EpiP including c-MYC-WT/ΔMB0 or MYCL-WT/ΔMB0 10 and 21 days later. Mean ± SD values are shown. n = 3, ***p < 0.001 by ordinary one-way ANOVA and Dunnett’s test vs. c-MYC-WT. The number of cells was counted using a Cell Counter model R1 (OLYMPUS).