Figure 4.

CRYAB regulates the BMSCs osteogenic differentiation via the WNT signalling. A, The expression of GSK‐3β, p‐GSK‐3β (Ser9), β‐catenin, p‐β‐catenin (Ser33/37/Thr41), LEF1 and AXIN2 at protein level in sh‐CRYAB group and sh‐Control group was measured by Western blotting. B, The expression of CTNNB1 at mRNA level in sh‐CRYAB group and sh‐Control group was measured by qRT‐PCR. C, The expression of GSK‐3β, p‐GSK‐3β (Ser9), β‐catenin, p‐β‐catenin (Ser33/37/Thr41), LEF1 and AXIN2 at protein level in CRYAB group and Vector group was measured by Western blotting. D, The expression of CTNNB1 at mRNA level in CRYAB group and Vector group was measured by qRT‐PCR. E, The expression of nuclear β‐catenin at protein level in sh‐CRYAB group and sh‐Control group was measured by Western blotting. F, Relative TOPFlash luciferase activity was measured in sh‐CRYAB group and sh‐Control group. G, The expression of nuclear β‐catenin at protein level in CRYAB group and Vector group was measured by Western blotting. H, Relative TOPFlash luciferase activity was measured in CRYAB group and Vector group. I‐L, The expression of AXIN2 and LEF1 at mRNA level was measured by qRT‐PCR. M, AR staining and ALP staining showed that the restricted osteogenic capacity of BMSCs resulting from the downregulation of CRYAB expression was rescued by the treatment with Wnt3a. N, AR staining and ALP staining showed that the enhanced osteogenic capacity of BMSCs resulting from the upregulation of CRYAB expression was reduced by the treatment with ICG‐001