Fig. 2: TGF-β1 enhances expression of Cx43 and pannexin1 of osteocytes.

a Gene profile of pannexins in osteocytes (MLO-Y4 cell line). Quantitative real-time PCR showed that pannexin1 was detected in osteocytes, whereas pannexin2 and pannexin3 were hardly detected. The results shown are based on three independent experiments (n = 3). b Gene profile of connexins in osteocytes (MLO-Y4 cell line). Quantitative real-time PCR confirmed that gene expression of Cx43 were much higher than other connexins, such as Cx44, Cx26, Cx30, and Cx31. The results shown are based on three independent experiments (n = 3). c Western blots of protein lysates from osteocytes (MLO-Y4 cell line) reveals the enhanced expression of Cx43 and pannexin1 in osteocytes after treatment with TGF-β1 (0.1, 1, 5, 10 ng/ml). d Quantitative analysis was performed to confirm the protein changes in c. The results shown are based on three independent experiments (n = 3). Data are presented as the mean ± s.d.; *p < 0.05 by t-test. e Western blots of protein lysates from osteocytes (MLO-Y4 cell line) reveal reduced expressions of Cx43 and pannexin1 in osteocytes after treatment with Repsox (25, 50 μM). f Quantitative analysis was performed to confirm the protein changes in e. The results shown are based on three independent experiments (n = 3). Data are presented as the mean ± s.d.; *p < 0.05 by t-test. g Representative IF staining by CLSM showing the increased expression of pannexin1 in osteocytes in response to TGF-β1 (5 ng/ml) (cytoskeleton, green; panx1, red; nucleus, blue). The results shown are based on three independent experiments (n = 3). h Representative IF staining by CLSM showing the increased expression of Cx43 in osteocytes in response to TGF-β1 (5 ng/ml) (cytoskeleton, green; Cx43, red; nucleus, blue). The results shown are based on three independent experiments (n = 3)