Fig. 3.

Preconditioning with KCs-CM further enhances DP signature recovery in 3D culture conditions. DP spheroids derived from cells previously cultured with KCs-CM had a significantly (a) lower DNA content, (b) higher amount of active ALP and (c) higher protein content (n = 3). Immunolabelling of DP spheroids preconditioned with KCs-CM showing (d) the proliferation-associated marker Ki67 (arrowheads) and the expression of (e) ALP, (f) α-SMA, (g) fibronectin, (h) collagen type I, (i) V1-isoform of versican, (j) V2-isoform of versican and the (k) gap junction protein connexin 43. Negative control is depicted on (l) and DAPI was used as the nuclear counterstaining (merged images, boxes). Data shown are mean ± s.e.m. *p < 0.05; **p < 0.01; ****p < 0.0001. Scale bars = 50 μm.