Skip to main content
. 2021 Feb 26;12:1293. doi: 10.1038/s41467-021-21545-1

Fig. 2. Postn promotes TSPC stemness and tenogenic differentiation potentials in early passage in vitro.

Fig. 2

a Immunofluorescence staining (i) and semi-quantification (ii) of Ki67 in PBS- and rPOSTN-treated TSPCs (n = 5 biologically independent samples, by two-tailed Student’s t test: ***P < 0.001). b (i) CFU-F assay of PBS- and rPOSTN-treated TSPCs. (ii) Semi-quantification of (i) (n = 4 biologically independent samples, by two-tailed Student’s t test: **P < 0.01). c Western blotting of Sox2 and Oct4 protein expression in TSPCs after rPOSTN treatment at different concentrations. d (i) CFU-F assay for assessing the anti-oxidative stress capacity of PBS- and rPOSTN-treated TSPCs under exposure to H2O2. (ii) Semi-quantification of (i) (n = 4 biologically independent samples, by two-tailed Student’s t test: **P < 0.01). e (i) SAβ-gal staining (top panel) and immunofluorescence staining of DNA injury-related protein γ-H2AX (bottom panel) of PBS- and rPOSTN-treated TSPCs suffering from H2O2 stimulation. Blank: without H2O2. The blue cells are senescent cells. (ii) Semi-quantification of (i) (n = 4 biologically independent samples, by one-way ANOVA with Tukey’s post hoc test: ***P < 0.001). f Western blotting of senescence-related protein P53, P21, and γ-H2AX of PBS- and rPOSTN-treated TSPCs suffering from H2O2 stimulation. g (i) Sirius Red staining (left panel) and Masson’s trichrome staining (right panel) of PBS- and rPOSTN-treated TSPCs. The pink and mazarine areas are positively stained respectively. (ii) Semi-quantification of (i) (n = 4 biologically independent samples, by one-way ANOVA with Tukey’s post hoc test: **P < 0.01, *P < 0.05). h Immunofluorescence staining (i) and semi-quantification (ii) of tenogenic markers Tnc, Col1, Tnmd, and Mkx in PBS- and rPOSTN-treated TSPCs (n = 3 biologically independent samples, by one-way ANOVA with Tukey’s post hoc test: ***P < 0.001, **P < 0.01, *P < 0.05). i (i) CFU-F assay of si NC- and si Postn-treated TSPCs. Si NC: negative control siRNA. (ii) Semi-quantification of (i) (n = 4 biologically independent samples, by two-tailed Student’s t test: **P < 0.01). j (i) Immunofluorescence staining of Ki67 of si NC- and si Postn-treated TSPCs. (ii) Semi-quantification of (i) (n = 4 biologically independent samples, by two-tailed Student’s t test: **P < 0.01). k (i) Representative images of SAβ-gal staining of si NC- and si Postn-treated TSPCs suffering from H2O2 stimulation. The blue cells are senescence cells. (ii) Semi-quantification of (i) (n = 4 biologically independent samples, by two-tailed Student’s t test: ***P < 0.001). l (i) Sirius Red staining (left panel) and Masson’s trichrome staining (right panel) of si NC- and si Postn-treated TSPCs (n = 4 biologically independent samples, by two-tailed Student’s t test: **P < 0.01). m (i) Immunofluorescence staining of tenogenic markers Tnc, Col1, Tnmd, and Mkx in si NC- and si Postn-treated TSPCs. (ii) Semi-quantification of (i) (n = 3 biologically independent samples, by two-tailed Student’s t test: **P < 0.01, *P < 0.05). Data are represented as mean ± SD. Exact P values were given in the Source Data file.