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. 2022 Dec 26;13:7952. doi: 10.1038/s41467-022-35687-3

Fig. 4. C6ST-1 overexpression potentiates mouse osteoblast differentiation.

Fig. 4

a The 4S/6S ratio of CS chains from parental, mock-transfected (Mock), or C6ST-1 overexpressing (C6ST-1 OE #1 and #2) MC3T3-E1 cells (n = 3 independent experiments, Dunnett’s multiple comparison test, two-sided). b ALP staining and mRNA expression for ALP (Akp2) in 14-day cultures of parental, mock, or C6ST-1 OE cells (#1 and #2) (n = 3 independent experiments, Dunnett’s multiple comparison test, two-sided). c Mineralized nodule formation in 21-day cultures of parental, mock, or C6ST-1 OE cells (#1 and #2) was assessed by Alizarin red staining. Data are obtained from three independent experiments and representative images are shown. (d) Adhesion of intact, ChABC-pretreated, or EDTA-treated MC3T3-E1 cells (parental or C6ST-1 OE #1 and #2) to plates precoated with recombinant N-cadherin or cadherin-11 (n = 10 fields from 3 independent experiments, for each condition, Tukey–Kramer multiple comparison method). e CS-C modulated the cadherin-mediated intracellular signaling pathways, including ERK1/2, Smad3, and Smad1/5/8, and upregulated Akp2 expression in semi-confluent, parental MC3T3-E1 cultures in the presence of an isotype control antibody (control Ab). Pretreatment with neutralizing antibodies for N-cadherin and cadherin-11 (cadherin Ab) abolished the CS-C effects (n = 3 independent experiments, Dunnett’s multiple comparison test, two-sided). Data in a, b, d, and e are represented as the mean ± s.d. Source data are provided as a Source Data file.