Fig. 5: Vldlr+ TSPCs demonstrated superior tenogenic differentiation capacity. | Nature Communications

Fig. 5: Vldlr+ TSPCs demonstrated superior tenogenic differentiation capacity.

From: Lymphatic–stem cell crosstalk promotes tendon regeneration via Notch1–Srebp2–mediated cholesterol metabolism

Fig. 5: Vldlr+ TSPCs demonstrated superior tenogenic differentiation capacity.The alternative text for this image may have been generated using AI.

A The expression of Vldlr and Lrp8 genes in TSCs, TPCs and Tenocytes. The center line denoted the median; the upper and lower bounds of the box represented the 25th and 75th percentiles; the whiskers referred to the data points within 1.5 times the interquartile range from the box. Statistical significance was analyzed by One-way ANOVA with Tukey’s post hoc test using R software. B The RNA expression of Vldlr in Neonate and Adult groups, analyzed by RT-qPCR analysis. C The expression of VLDLR in Neonate and Adult groups, assessed by western blotting. D Confocal imaging of tendon sections from Vldlr-CreERT2, tdTomato mice that had been induced by tamoxifen at P3 of age. Mice were analyzed at 7 days, 1 months and 3 months after the induction. E Flow cytometry analysis of enzymatically dissociated tendon cells from Vldlr-CreERT2, tdTomato mice that had been induced by tamoxifen at P3; Col1α1 were stained to indicate the tendon cells. F Confocal imaging of tendon sections from Vldlr-CreERT2, tdTomato mice that had been induced by tamoxifen at 1 month of age. Mice were analyzed at 7 days, 1months and 3 months after the induction. G Flow cytometry analysis of enzymatically dissociated tendon cells from Vldlr-CreERT2, tdTomato mice that had induced by tamoxifen at 1 month; Col1α1 were stained to indicate the tendon cells. H Images of renewed Vldlr- tdTomato positive cells and FMOD expression in 30 d regenerated tendons. I The proliferation of Vldlr + and Vldlr– TSPCs, analyzed by Ki67 staining. J The Sirius red, Masson trichrome staining and semi-quantification in Vldlr+ and Vldlr– TSPCs. K The HE staining, Masson trichrome staining and collagen content analysis of regenerated tendon in control, defects, Vldlr + and Vldlr – TSPCs transplant groups. L The expression of VLDLR and Ki-67 in the control and injured Relnfl/fl and Relncko tendons, analyzed by immunofluorescence. Each dot represents one individual replicate (n = 3 biological replicates in B, I, J； n = 5 biological replicates in E, G, K). Statistical significance was analyzed by One-way ANOVA with Tukey’s multiple comparisons test (A, E, G, K). Statistical significance was analyzed by unpaired Student’s t test (B, I, J).

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