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Fig. 5 | Stem Cell Research & Therapy

Fig. 5

From: Dextran sulfate prevents excess aggregation of human pluripotent stem cells in 3D culture by inhibiting ICAM1 expression coupled with down-regulating E-cadherin through activating the Wnt signaling pathway

Fig. 5

Interfering ICAM1 endogenously by lentivirus-mediated shRNA. A The EGFP+ colonies after lentivirus transduction and puromycin selection. Representative images were shown. Scale bar = 200 μm. B Gene expression analysis by qRT-PCR for ICAM1 in the H9 cells transduced with the lentivirus expressing different shRNAs, relative gene expression represents data normalized to GADPH and expressed relative to cells transduced with the lentivirus expressing shNT. C The EGFP+ aggregates were shown in suspension culture on day 5. Representative images were shown. Scale bar = 200 μm. D Comparison of average diameter of the aggregates of the H9 cells in two groups. E Gene expression analysis by qRT-PCR for ICAM1 and OCT4 in the aggregates transduced with the lentivirus expressing shRNA-b and shNT, respectively, relative gene expression represents data normalized to GADPH and expressed relative to cells transfected by shNT. (F) The protein levels of E-cad and ICAM1 were determined by western blotting in the aggregates transduced with the lentivirus expressing shRNA-b and shNT, respectively, in two groups. G The densitometry for the protein levels of ICAM1 was quantitated in F; GAPDH was used as a loading control. Data represent the mean ± SD. *P < 0.05 and **P < 0.01

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Stem Cell Research & Therapy

ISSN: 1757-6512