Supplementary Figure 7: Characterization of iPSC clones.
From: Autophagy and mTORC1 regulate the stochastic phase of somatic cell reprogramming
(a) Genomic PCR analysis of transgene integration of 3 representative iPSC clones generated with retroviral 4F and shBeclin1, shVps34 and Atg16l-M. Negative indicates ESCs; positive indicates Actin. (b) Phase contrast, fluorescence and immunofluorescence microscopy of representative iPSC clones. Scale bars: 100 μm. (c) Bright-field view of Atg5 KO 4F iPSC clone C1. Scale bar: 100 μm. (d) Western blotting for LC3B and ATG5-12 in Atg5 KO-4F C1. (e) qPCR analysis of the expression of endogenous pluripotent genes and the silencing of 4F transgenes in iPSCs. Data are plotted from 1 experiment with 3 technical replicates. (f) DNA methylation profile of the Oct4 and Nanog proximal promoters in the indicated iPSCs, ESCs and MEFs. (g) Karyotype analysis of these iPSCs. (h) Hematoxylin/eosin staining of the teratomas formed from the shRNA iPSCs. Scale bar: 100 μm. (i) Chimeric mice produced with the above clones and germline transmission using Atg16l-M-C2 iPSCs (F1). Source data are provided in Supplementary Table 4. Uncropped images of blots are shown in Supplementary Fig. 6.
