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. 2016 Nov 18;2(11):e1600874. doi: 10.1126/sciadv.1600874

Fig. 5. Nestin is important for self-renewal and phenotypic maintenance of TSPCs.

Fig. 5

(A) Western blot and qPCR demonstrated successful knockdown of nestin expression in TSPCs. (B) CFU assay showed that self-renewal of TSPCs was decreased by the knockdown of nestin (n = 3). (C) Immunofluorescence staining of F-actin, expression in TSPCs infected with RNA interference (RNAi) against nestin (shRNA-Nes/shRNA-Nes1), or a control scrambled sequence (mock). (D) Quantitative data on cell morphology change of TSPCs under nestin knockdown. Results are presented as means ± SD (n = 40 to 60). Scale bars, 50 μm. (E) qPCR analyses showing that the expression of tendon-associated genes in TSPC cell sheets from the control group was significantly higher than that from the nestin knockdown group (n = 5). (F) Transmission electron micrographs of collagen fibrils in cell sheets from the control and nestin knockdown groups. *P < 0.05, **P < 0.01 (one-way ANOVA). Results are presented as means ± SD (n = 5). Scale bars, 50 μm (B); 200 nm (D).