Figure 6

Synemin knockdown in mouse ES cells by shRNA lentiviral particles. A, Analysis of mRNA expression by qRT-PCR. Messenger RNA was extracted from ES cells infected by control or synemin shRNA lentivirus (shSyn). The PCR analysis was performed with SYBR green PCR technology. HPRT was used as a control gene for normalization. B, Western blotting analysis of synemins M (Syn M), Keratin 8 (K8) and Oct-3/4 proteins extracted from ES cells infected by control or synemin shRNA lentivirus (shSyn). GAPDH was used for the loading control. C-N, Immunochemistry analysis demonstrating ES cells colonies labeled for Oct-3/4 (C, F), keratin 8 (I, L) and synemin M (D, G, J, M) in mouse ES cells infected by control (C-E, I-K) or synemin shRNA lentivirus (shSyn, F-H, L-N). Synemin knockdown in mouse ES cells by shRNA lentivirus has no influence on expression of Oct4, but decrease keratin 8 expression. Bar = 20 μm.