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. 2021 Feb 25;16(3):610–625. doi: 10.1016/j.stemcr.2021.01.014

Figure 6.

Figure 6

iPSC-derived hypertrophic chondrocytes allow drug testing and further exploration of chondrodysplasia pathology

(A) Number of differentially expressed genes by microarray analysis using FDR-adjusted or unadjusted p value of <0.05 by moderated t test in each mutant compared with its isogenic control.

(B and C) Top five Tissue Development (B) and Cellular Movement (C) terms in IPA Diseases and Functions for the MATN3 T120M and COL10A1 S600P mutants versus their isogenic controls by microarray analysis. Genes with adjusted p value of <0.05 by moderated t test were used for IPA. Red bars, positive activation Z score; blue bars, negative activation Z score; gray bars, zero or unavailable activation Z score.

(D) Heatmap of the log2 fold change of UPR genes from different branches in the COL10A1 S600P and MATN3 T120M mutants by microarray analysis.

(E and F) mRNA expression of ER stress markers (E) and chondrocyte marker COL2A1 (F) by qPCR in the MATN3 T120M mutant with and without drug treatment.

(G) Pellet size of the MATN3 T120M mutant with and without treatment.

(H) ER size of the MATN3 T120M mutant with and without treatment, quantified from the area of PDI fluorescence.

(I) Intracellular retention of MATN3 in the MATN3 T120M mutant with and without treatment, quantified from fluorescence intensity of MATN3 co-staining with PDI (intracellular) divided by total fluorescence intensity.

(J) Immunostaining of MATN3 and PDI in the MATN3 T120M mutant with and without treatment. Arrowheads indicate intracellular aggregates. Similar results were obtained in n = 4 independent experiments. Scale bars, 100 μm.

All results are from day 56 of HI. The number of independent experiments is n = 3 in (A) to (D) and n = 4 in (E) to (I). In (E) to (I), values are expressed as mean ± SEM, relative to the isogenic control (dotted lines), and statistical analysis was performed by unpaired two-sided t test of treated compared with untreated samples (n.s., no significant difference; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001). As experiments were performed at the same time, data of untreated mutants and isogenic controls in (E) to (J) are the same as in Figures 2G, 3D, 4E, 4F, and S5A. CBZ, carbamazepine; TMAO, trimethylamine N-oxide; VPA, valproic acid; RM, rapamycin. See also Figure S6 and Table S1.