Fig. 3: Molecular characterization of the GDF5 GROW1 enhancer and rs4911178 variant in human chondrocytes and the mouse model.
From: Joint disease-specificity at the regulatory base-pair level
a Expression of CEP250 (non-significant (0.082)), GDF5 (significant, p < 0.018), and UQCC1 (p = 0.099) in human T/C-28a2 chondrocytes lacking the GROW1 enhancer (n = 3 biologically independent samples). b Expression of CEP250 (p = 0.849), GDF5 (p = 0.011), and UQCC1 (p = 0.731) in T/C-28a2 cells lacking a 13 bp region containing rs4911178 in GROW1 (n = 3 biologically independent samples). c ASE assays on E15.5 proximal (p = 0.019) and distal femur (p = 0.038) chondrocytes from C57BL/6 J/129SvJ GROW1rs4911178-A/rs4911178-+ replacement mice (n = 7 biologically independent samples). d PITX1 ChIP assay on a 234 bp GROW1 sub-region containing rs4911178 in human T/C-28a2 chondrocytes showing input (left) and pull-down (right) in the gel image. This experiment was repeated three times independently with similar results. e PITX1 ChIP assay on a 255 bp GROW1 sub-region containing rs4911178 in GROW1rs4911178-A/rs4911178-G mice showing input and pull-down from E15.5 proximal femur. This experiment was repeated three times independently with similar results. f A/G allelic ratio in PITX1 ChIP pull-down compared to input from GROW1rs4911178-A/rs4911178-G mice (39.6% decrease in ‘A’ over G, p = 0.002 in the proximal femur; 34.5% decrease in ‘A’ over ‘G’, p = 0.002 in distal femur; 20.2% decrease in ‘A’ over ‘G’, p = 0.001 in proximal tibia) (n = 3 biologically independent samples). g Fold-change for PITX1 and Gdf5 gene expression in PITX1 over-expressed chondrocytes cultured from E15.5 embryonic hind limb buds (proximal femoral, distal femoral, and proximal tibia). Summary data are presented as mean and standard deviations. All p-values are two-sided.
