Figure 6.
Different Mgarp Expression Levels Required at Early and Late Stages during Reprogramming of Somatic Cells into iPSCs Are Competitively Regulated by OCT4 and KLF4
(A) Reprogramming of MEFs, NSCs, and BMCs into iPSCs using different factor combinations. Representative wells were stained for ALP activity (red) on day 21 after infection.
(B) Oct4-GFP-positive iPSC colonies were counted on day 21 after infection with different factor combinations. The error bars represent SD of the mean (biological replicates, n = 5). p values refer to a two-tailed, unpaired Student’s t test: ∗p < 10−3; ∗∗p < 10−4; ∗∗∗p < 10−5.
(C) Monitoring of Mgarp expression levels (qRT-PCR) during a 15-day period after infection of MEFs with different combinations of genes, normalized to ESCs. The error bars represent SD of the mean (biological replicates, n = 4).
(D) Oct4-GFP-positive iPSC generated after infection of MEFs with four factors plus tetracycline-inducible (Tet-) vectors for Mgarp overexpression or knockdown. The expression or inhibition was induced at different time points, as indicated. The error bars represent SD of the mean (biological replicates, n = 5). p values refer to a two-tailed, unpaired Student’s t test: ∗p < 10−3; ∗∗p < 10−4; ∗∗∗p < 10−5; n.s., not significant (p > 0.1). Abbreviations: shRNA, small hairpin RNA; OSKM, Oct4, Sox2, Klf4, and Myc.
See also Figures S4–S7 and Table S5.