Extended Data Fig. 2: Bcl11b constrains reprogramming & transformation.
(a) Alkaline Phosphatase (AP) staining of iPS colonies. One representative experiment (from three independent experiments). (b) Counting of AP-positive colonies depicted in (h). Data are the mean ± s.d. (n = 3 independent experiments). Two-tailed student’s t-test was used. (c) Foci staining generated from Thy1low and Thy1high cells FACS-sorted at day 5 of Transfo. One representative experiment (from three independent experiments). (d) Counting of foci depicted in (c). Data are the mean ± s.d. (n = 3 independent experiments). Two-tailed student’s t-test. (e) Relative transcript level of Bcl11b. Data are the mean ± s.d. (n = 4 independent experiments for left panel and 3 for right panel). (f) Bcl11b expression profiles during MEFs and keratinocytes reprogramming. Data are extracted from Nefzger et al., 2017. (g-h) Bcl11b knockdown efficiency. (g) Q-RTPCR of Bcl11b levels 3 days after infection. Data are the mean + /- sd of 3 independent experiments. Two-tailed student’s t-test. (h) Western blot for Bcl11b in similar settings as (g). (i) Representative pictures of foci. (j) Counting of foci depicted in (i). Data are the mean ± s.d. (n = 3 independent experiments). Two-tailed Student’s t-test. (k) Pictures of AP + iPS colonies generated from Bcl11b conditional KO MEFs treated or not with Tamoxifen. (l) Countings of colonies depicted in (k). Data are the mean ± s.d. (n = 8 independent experiments). Two-tailed student’s t-test. (m) Patterns of MEF identity signature score using gene lists from Nefzger et al., 2017 and Schiebinger et al., 2019. (n) Western blot of Bcl11b in the tdTomato-high/low fractions. (o) FACS analysis of Bc11b-tdTomato. (p) Alkaline phosphatase (AP) staining of iPS colonies generated from tdTomatolow and tdTomatohigh cells. One representative experiment (from four independent experiments). (q) Counting of AP-positive colonies depicted in (p). n = 4 independent experiments. Two-tailed Student’s t-test. (r) Foci staining of cells generated from tdTomatolow and tdTomatohigh cells. One representative experiment (from three independent experiments). (s) Counting of foci depicted in (r). Data are the mean ± s.d. (n = 3 independent experiments). Two-tailed Student’s t-test.
