Skip to main content
Fig. 5 | BMC Biology

Fig. 5

From: Nucleus size and DNA accessibility are linked to the regulation of paraspeckle formation in cellular differentiation

Fig. 5

NEAT1_2 depletion increases differentiation potential. a The strategy of generating NEAT1−/−, NEAT1STOP, and NEAT1ΔTH hESC clones by CRISPR/Cas9. b–d Representative images (b) and quantification of paraspeckles by smFISH with NEAT1_2 (red) probe (c) or RT-qPCR (d) in parental (WT), NEAT1ΔTH, NEAT1−/−, and NEAT1STOP hESCs after 3 days of differentiation induced by retinoic acid (b, c) or spontaneous differentiation (d). Primers for total NEAT1 targeted both NEAT1 isoforms. DAPI staining in blue; scale bar: 10 μm. Low levels of paraspeckles were assigned to NEAT1−/− and NEAT1STOP hESCs due to background signals. e–i RT-qPCR of pluripotency and differentiation markers (e), and flow cytometry of pluripotency markers after WT, NEAT1ΔTH and NEAT1−/− hESCs were spontaneously differentiated for 3 days (f, g) or after 4 days of neuroectoderm differentiation (h, i). NEAT1STOP hESCs were included in e. n (# of experiments / # of clones) = 2/2 in d, 2/3 in e, g (except NEAT1STOP which is 3/2) and 1/3 in i. Error bars represent standard deviation in ce or standard error of the mean in g, h. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, unpaired t-test

Back to article page