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. 2019 Aug 28;5(8):eaax2476. doi: 10.1126/sciadv.aax2476

Fig. 4. Effects of morphogen priming of engineered mesenchymal condensations on in vitro chondrogenic lineage specification at the time of implantation.

Fig. 4

Histological Safranin-O/Fast green staining of representative microparticle-containing hMSC sheets at the time of implantation (2 days; n = 3 per group). Scale bars, 100 μm (top, x10; bottom, x40 magnification of dotted squares). (B) Normalized mRNA fold change over control of key chondrogenic or osteogenic markers by quantitative reverse transcription polymerase chain reaction (qRT-PCR; n = 3 per group; *P < 0.05, **P < 0.01, ***P < 0.001 versus empty/control; $P < 0.05 versus BMP-2–containing hMSC sheets). (C) Representative immunoblots and (D) relative quantification of phosphorylated SMAD5 (p-SMAD5)/SMAD5 and (E) p-SMAD3/SMAD3 in lysates of day 2 hMSC sheets (n = 3 per group). β-Actin served as the loading control. Individual data points are shown as means ± SD. Comparisons between groups were evaluated by one-way ANOVA with Tukey’s post hoc tests. Repeated significance indicator letters (a, b, and c) signify P > 0.05, while groups with distinct indicators signify P < 0.05.