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. 2023 Jan 19;12:e77257. doi: 10.7554/eLife.77257

Figure 3. ECM defects in Ddr2-deficient synchondroses.

(a-d) Immunofluorescent staining of ISS and SOS sections from 2-week-old WT and Ddr2slie/slie synchondroses. (a) Representative images of COL2 immunostaining (red) in ISS and SOS showing homogenous distribution around chondrocytes in WT synchondroses, while Ddr2slie/slie mice showed uneven, ring-like immunostaining around chondrocytes (white arrowheads). Boxed region is shown in higher magnification (bottom). Scale bar: 200 μm. (b) Immunofluorescent images of COL10 immunostaining in the hypertrophic zone (HZ) of synchondroses showing no major changes in staining distribution between WT and Ddr2slie/slie mice. Scale bar: 50 μm. (c) Immunofluorescence images showing COL1 (red) staining in trabecular (arrowheads) and cortical (arrows) bones of the cranial base in WT and Ddr2slie/slie mice. Scale bar: 200 μm. (d) Immunofluorescent images showing IBSP (red) in trabecular (arrowheads) and cortical (arrows) bones of cranial base is decreased in Ddr2slie/slie synchondrosis compared with WT littermates. See Figure 3—figure supplement 2 for quantitation of IF staining. Scale bar: 200 μm. Cell nuclei were stained with DAPI (blue) in a–d. Bm: Bone marrow; Br: Brain; Tb: Trabecular bone; Ct: Cortical bone; Ms: Muscle.

Figure 3.

Figure 3—figure supplement 1. Ddr2 deficiency is associated with loss of collagen organization in the ISS of Ddr2slie/slie mice.

Figure 3—figure supplement 1.

(a) Second harmonic generation imaging of fibrillar collagen in the ISS resting zone of 2-week-old WT versus Ddr2slie/slie mice. Scale bar, 100 µm (b) anisotropy was measured using ImageJ FibrilTool as described in Methods. For each sample, 3 or 4 ROIs were selected within the central ISS resting zone for analysis and averaged (n=3 or 4/group). Bars are mean +/-SEM.
Figure 3—figure supplement 2. Quantitation of Ibsp (Bsp) and Col I immunofluorescence in WT and Ddr2slie/slie mice.

Figure 3—figure supplement 2.

(a) Ibsp in hypertrophic chondrocyte layers. (b) Ibsp in primary spongiosa. Note: Ibsp is significantly lower only in the SOS. (c) No change in Col I in the primary spongiosa. Data are presented as mean ± SD. *p<0.05.