Fig. 6: Osteogenic differentiation in fibroblasts from the patient and controls.
From: Enhancer hijacking at the ARHGAP36 locus is associated with connective tissue to bone transformation
a Schematic representation of the osteogenic protocol. Cells (n = 1 proband, n = 3 controls; 3 technical replicates of each sample) were stimulated with glycerophosphate (10 mM), ascorbic acid (50 µM), and dexamethasone (100 nM), were collected in six-time points (0–5 weeks) and stained for alizarin red (AR) for calcium deposition. b Patient samples showed a faster osteogenic differentiation after two weeks of stimulation, lasting 3-4 weeks. The control values at each time point were set to 1.0. The translucent band shows the confidence interval for each replicate for the respective time point. c Schematic representation of HH inhibition by GANT61 (1 and 5 µM) during the osteogenic differentiation. d GANT61 (5 µM) reduces by half GLI1 expression in both patient and control samples after 48 h of molecule exposure (n = 1 proband, n = 1 control). Pb: proband; Ct: controls. RT-qPCR quantification is expressed as mean fold ±SD in arbitrary units. Source data are provided as a Source Data file. e HH inhibition by GANT61 (5 µM) revealed a minor decrease of 10% of AR staining in all samples. The median score is represented by the horizontal line in the center. The 25th and 75th percentile values are indicated by the lower and upper limits of the box. Source data are provided as a Source Data file.
