Research Paper Volume 12, Issue 8 pp 7431—7447

Sirt1 is regulated by miR-135a and involved in DNA damage repair during mouse cellular reprogramming

Figure 1. miR-135a impeded reprogramming efficiency partly through inhibiting Sirt1. (A) The effects of miR-135a precursor or inhibitor on relative number of iPSC colonies formed from 1° (n=4) or 2° (n=7) MEFs on D10/15 after Doxycycline (DOX) induction. miRNA precursor (pre-ctrl) and inhibitor (inh-ctrl) with scrambled sequence were used as controls (*:p<0.05; t-test). (B) The relative expression levels of miR-135a in MEFs and mESCs (n=3; *: p<0.05; t-test). (C) The relative expression levels of miR-135a in 1° MEFs after transfecting with miR-135a precursor (pre-135a) or inhibitor (inh-135a) for 72h (n=4; *:p<0.05; t-test). (D) The relative SIRT1 and SIRT6 protein levels in 1° MEFs after transfecting with miR-135a precursor (pre-135a) or inhibitor (inh-135a) for 72h (n=4; *:p<0.05; t-test). (E) The relative expression levels of miR-135a in mESCs and EBs at day 17 (EB-D17, n=3; #:p<0.001; t-test). (F) The relative miR-135a expression levels during reprogramming of 2° MEFs from day 0 to day 21 (n=3; *:p<0.05; t-test). (G) The relative expression levels of miR-135a in 1° MEFs after RSV treatment for 72h (n=4; t-test).