Fig. 2

EMT of HPVC cells. a After 6 days of FGF8/FGF2/VEGF treatment on MEFs, valve progenitors (HPVCs) were recovered with trypsin, seeded on fibronectin-coated wells and treated with 100 ng/ml BMP2. After 2 days, RNA was recovered and cDNAs were run in real-time PCR for post EMT markers. BMP2 samples are normalized on control (before treatment) samples, showing an increase in the expression of post-EMT markers Data are representative of 5 cell differentiation and EMT- induction experiments.Boxes and whiskers (min to max) show the values lower than the 2.5th percentile and greater than the 97.5th percentile as circles. (*) significantly different p ≤ 0.01. b t-distributed stochastic neighbor embedding (t-SNE) 2D cell map 10X genomics (n = 3700) cells (left panel). Highlight of cell populations expressing TAGLN and genes marking more specifically fibrosa (COL1A1) or spongiosa (CD9, TDGF1) (middle panel) and heatmap of graph-based Log2 fold changes in gene expression of cell cluster compared to all other cells (right panel). c HPVCs give rise to tendinous/chondrogenic cells. HPVCs were pelleted into a tube and treated with a chondrogenic medium as described in the methods. Real-Time PCR shows an upregulation of Scleraxis and Collagen 1a (Col 1) genes versus non-treated cells following three weeks of treatment (two experiments in duplicate; (*) significantly different p ≤ 0.01). d The cell pellets were embedded in paraffin and 5 um sections stained with anti Sox9, anti-Collagen1a and anti-calcitonin antibodies. Data are representative of 3 experiments. The scale bar indicates 50 μm. Source data are provided as a Source Data file