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. 2020 Nov 17;11(11):1360. doi: 10.3390/genes11111360

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Hedgehog signaling pathway inhibitor, Cyclopamine, blocked the inhibitory effect of 20S on OA-induced adipogenic differentiation of chicken-bone marrow stem cells (cBMSCs). When confluent, cBMSCs were pretreated for 2 h with control vehicle (VEH) or 4 µM cyclopamine (CYC) followed by 24 h and 48 h of treatment with (1) control vehicle (C), (2) Oleic acid (OA), (3) OA + 10 µM 20S, or (4) 10 µM 20S. RNA was isolated, reverse transcribed to cDNA, and q RT-PCR was conducted to analyze osteogenic gene expression. GAPDH was used as a housekeeping gene. Fold changes in gene expression relative to the control were calculated using the −ΔΔCt method. mRNA expression of (A1,A2) FABP4, (B1,B2) PPARγ, (C1,C2) c/EBPα, (D1,D2) c/EBPβ, and (E1,E2) LPL are presented above. Data from the representative experiment are reported as the mean of the triplicate determination. Bars represent mean ± SEM for triplicate determinations. Bars without a letter (a–b) in common are significantly different (p < 0.05) when analyzed with Tukey’s test.

Hedgehog signaling pathway inhibitor, Cyclopamine, blocked the inhibitory effect of 20S on OA-induced adipogenic differentiation of chicken-bone marrow stem cells (cBMSCs). When confluent, cBMSCs were pretreated for 2 h with control vehicle (VEH) or 4 µM cyclopamine (CYC) followed by 24 h and 48 h of treatment with (1) control vehicle (C), (2) Oleic acid (OA), (3) OA + 10 µM 20S, or (4) 10 µM 20S. RNA was isolated, reverse transcribed to cDNA, and q RT-PCR was conducted to analyze osteogenic gene expression. GAPDH was used as a housekeeping gene. Fold changes in gene expression relative to the control were calculated using the −ΔΔCt method. mRNA expression of (A1,A2) FABP4, (B1,B2) PPARγ, (C1,C2) c/EBPα, (D1,D2) c/EBPβ, and (E1,E2) LPL are presented above. Data from the representative experiment are reported as the mean of the triplicate determination. Bars represent mean ± SEM for triplicate determinations. Bars without a letter (a–b) in common are significantly different (p < 0.05) when analyzed with Tukey’s test.