Additional file 4 of Nucleus size and DNA accessibility are linked to the regulation of paraspeckle formation in cellular differentiation

Figure S4. Related to Fig. 5, characterization of NEAT1-manipulated cells. a RT-qPCR of pluripotency and differentiation markers of undifferentiated NEAT1−/−, NEAT1STOP and NEAT1ΔTH hESC clones. b, c Flow cytometry analysis of pluripotency surface markers TRA1–60 and SSEA5 after 2 days of spontaneous differentiation of WT, NEAT1ΔTH and NEAT1−/− hESCs. d RT-qPCR time course analysis of pluripotency and neural marker genes during differentiation towards neural rosettes which appeared around day 12 of the differentiation towards NSCs. Same cell lines as in b, c. e-g RT-qPCR analysis of NEAT1ΔpA hESC clones differentiated to lateral mesoderm (e), definitive endoderm (f) and neuroectoderm by 4 days differentiation of NSCs (g). h-k Representative histograms and quantification of flow cytometry analysis for pluripotency markers in pluripotent (h, j) NEAT1STOP hESCs and after 3 days of spontaneous differentiation (i, k). Forward and side scatter gating was employed to gate out debris and cell clumps. n (# of experiments / # of clones) = 3/2 in a, 1/3 in c, e, f, 2/3 in d, g and 2/2 in j, k. Error bars represent standard deviation.