Fig. 5

ANA promotes bone development by enhancing bone formation through butyrate. A–C Measurement of the body weight of chicks (A), content of short-chain fatty acids (SCFAs) in cecal contents (B), and relative mRNA expression in the femur (C) after 14 days of inoculation. n = 7–8 per group. D Schematic representation of the experiment in vitro experiment used to investigate the effect of ANA fermentation products and NaB on osteogenesis in primary osteoblasts E The mRNA expression of runt-related transcription factor 2 (Runx2), osteocalcin (OCN), and bone morphogenetic protein 2 (BMP2) after supplementation with 0.01%, 0.1%, and 1% ANA fermentation products for 3 days. n = 3 per group. F ALP activity after supplementation with 0.01%, 0.1%, and 1% ANA fermentation products for 3 days. n = 3 per group. G Representative images and quantification of ARS staining after supplementation with 0.01%, 0.1%, and 1% ANA fermentation products for 9 days. n = 6 per group. H The mRNA expression of Runx2 , OCN, and BMP2 after 10 −6 M, 10 −5 M, or 10 −4 M NaB supplementation for 3 days. n = 3 per group. I ALP activity after 10 −6 M, 10 −5 M, or 10 −4 M NaB supplementation for 3 days. n = 3 per group. J Representative images and quantification of ARS staining after 10 −6 M, 10 −5 M, or 10 −4 M NaB supplementation for 9 days. n = 6 per group. K Schematic representation of the in vitro experiment used to investigate the effect of ANA or NaB on osteoclastogenesis in primary osteoclasts. n = 6 per group. L–M TRAP activity (L) and total protein concentration of osteoclasts (M) after administration of 1% ANA fermentation products or 10 −4 M NaB for 6 days. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001