Figure 2.

COL10A1 and MATN3 mutants show no disruption of chondrogenic differentiation

(A) Percentage of DLL1-positive cells on day 2 of SI.

(B) mRNA expression of the heterozygously mutated gene by qPCR, with values from a second clone also shown for each mutant and isogenic control (number of independent experiments shown in Table S2). Values are relative to the mean of six pieces of the human distal femoral growth plate. Statistical analysis by ANOVA and post hoc Tukey’s HSD.

(C–F) Safranin O staining (top panels) or COL2 (green) and COL1 (red) immunostaining (bottom panels) of mutants (left panels) and their isogenic controls (right panels). Similar results were obtained in n = 4 independent experiments. Scale bars, 100 μm.

(G) Pellet size of each mutant and its isogenic control. Three technical replicates per independent experiment were measured.

(H) Cell size of each mutant relative to the isogenic control, quantified from the inverse image of COL2 fluorescence.

(I) TUNEL-positive cells per FOV (field of view) relative to the isogenic control of each mutant.

All results except (A) are from day 56 of HI. Values are expressed as mean ± SEM. Dotted lines indicate the value = 1 of the isogenic controls in (H) and (I). Except where stated otherwise, the results are from n = 4 independent experiments, and statistical analysis was performed using unpaired two-sided t tests. (n.s., no significant difference; ^∗p < 0.05, ^∗∗p < 0.01). See also Figures S2–S5.