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. 2009 Mar 17;4(3):e4914. doi: 10.1371/journal.pone.0004914

Figure 3. Shp2 knockdown in hESCs leads to impaired differentiation.

Figure 3

(A) Immunoblotting and qRT-PCR indicate efficient Shp2 knockdown by 80% in hESCs. H14 cells were transfected with Shp2-specific (+siShp2) or non-specific siRNA (−siShp2), and cells were cultured on matrigel-coated dishes in CM for 2 days, total cell lysates were immunoblotted with the indicated antibodies or total RNAs were extracted for qRT-PCR (n = 3, ** P<0.01). (B) Immunostaining of NANOG, OCT3/4, SOX2 in hESCs. After transfection, H14 cells were cultured in CM for 2 days, then the medium were changed to UM to induce differentiation for additional 6 days. Scale bars, 10 µm. (C) Total RNAs were extracted from hESCs cultured in CM or UM, qRT-PCR was performed, and the relative values of Shp2 knockdown over control hESCs were indicated (n = 3, * P<0.05, ** P<0.01).