Fig. 2

Restriction is observed at the pluripotent cell state. a LV were applied to C57BL/6 p14f/f adult fibroblasts (Ad fib) (n = 8) or C3H Mefs (n = 3) at an MOI of 10 and 100 (independently produced viral supernatants). The percentage of EGFP positive cells was determined by flow cytometry. NTD non-transduced control. b Scheme of iPSC differentiation by withdrawal of LIF and feeder cell co-cultivation (upper panel). Representative fluorescence microscopy pictures of transduced differentiated cultures with LV.DsRed or GV.DsRed at an MOI of 100, SSEA1 live staining in green and an overlay of all channels are depicted (lower panel). c Differentiated cultures were transduced with LV (n = 5) and GV (n = 4) encoding EGFP at an MOI of 100 (independently produced viral supernatants). Pluripotent (SSEA1+) and differentiated (SSEA1−) cells were analyzed by flow cytometry. One-way ANOVA with Tukey-Kramer post hoc test was used for statistical analyses. * p = 0.049; ns p = 0.706. d ECC transduced with LV (n = 3) or GV (n = 2) at an MOI of 10 and 100 (independently produced viral supernatants). The percentage of EGFP positive cells was analyzed by flow cytometry