Journal: JBMR Plus

Article Title: Follistatin-like protein 1 (FSTL1) modulates bone remodeling and attenuates bone loss in a mouse model of postmenopausal osteoporosis

doi: 10.1093/jbmrpl/ziag037

Figure Lengend Snippet: Fstl1 enhances the osteogenic differentiation potential of MC3T3-E1 cells. (A) Representative alkaline-phosphatase (ALP) staining at 7 d. (B) Quantitative ALP activity. (C) Representative Alizarin Red S (ARS) staining at 14 d. (D) Quantification of ARS absorbance (OD 562 ). (E-H) Relative mRNA expression of Runx2, Sp7, Ocn, and Opn at 14 d determined by RT-qPCR. (I) Western-blot bands for RUNX2 and SP7. (J and K) Densitometric ratios of RUNX2/GAPDH and SP7/GAPDH. Sample sizes: con ( n = 3), OE-Fstl1 ( n = 3), and sh-Fstl1 ( n = 3). Data are presented as mean ± SD; inter-group comparisons were performed using 1-way ANOVA. * p < .05, ** p < .01, *** p < .001, **** p < .0001; ns, not significant.

Article Snippet: Antibodies against RUNX2, SP7, NFATC1, and CTSK were purchased from Abcam, while the anti-GAPDH antibody and anti-FSTL1 antibody were obtained from Proteintech.

Techniques: Staining, Activity Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: JBMR Plus

Article Title: Follistatin-like protein 1 (FSTL1) modulates bone remodeling and attenuates bone loss in a mouse model of postmenopausal osteoporosis

doi: 10.1093/jbmrpl/ziag037

Figure Lengend Snippet: Fstl1 suppresses the osteoclast-differentiation activity of BMMs. (A) TRAP staining showing TRAP + multinucleated osteoclast morphology in each group. (B) Quantitative analysis of TRAP + osteoclast numbers. (C) Dual staining of F-actin rings (green) and DAPI (blue) illustrating the resorptive synapse structure of mature osteoclasts. (D) Average number of nuclei per TRAP + osteoclast. (E and F) Representative images of resorption pits on an osteoassay surface from each group and quantification of the total resorbed area per well (percentage of surface area). (G-J) Relative mRNA expression of Ctsk, Nfatc1, c-Fos, and Trap determined by RT-qPCR. (K) Western-blot bands for CTSK and NFATc1. (L and M) Densitometric ratios of NFATc1/GAPDH and CTSK/GAPDH. Sample sizes: con ( n = 3), OE-Fstl1 ( n = 3), and sh-Fstl1 ( n = 3). Data are presented as mean ± SD; inter-group comparisons were performed with 1-way ANOVA. * p < .05, ** p < .01, *** p < .001, **** p < .0001; ns, not significant.

Article Snippet: Antibodies against RUNX2, SP7, NFATC1, and CTSK were purchased from Abcam, while the anti-GAPDH antibody and anti-FSTL1 antibody were obtained from Proteintech.

Techniques: Activity Assay, Staining, Expressing, Quantitative RT-PCR, Western Blot

Journal: JBMR Plus

Article Title: Follistatin-like protein 1 (FSTL1) modulates bone remodeling and attenuates bone loss in a mouse model of postmenopausal osteoporosis

doi: 10.1093/jbmrpl/ziag037

Figure Lengend Snippet: Age-associated down-regulation of FSTL1. (A) Serum FSTL1 concentrations in the normal and osteoporosis groups measured by ELISA. (B) Relative FSTL1 mRNA expression in peripheral blood leukocytes from the 2 groups determined by qPCR. (C) GTEx analysis of sex differences in blood-cell FSTL1 expression. (D) GTEx comparison of FSTL1 expression in individuals aged <50 yr and ≥50 yr. (E) GTEx comparison of FSTL1 expression in individuals aged <70 yr and ≥70 yr. (F) Age-related trend of blood-cell FSTL1 expression in men. (G) Age-related trend of blood-cell FSTL1 expression in women. Data are expressed as mean ± SD; 2-group comparisons were performed with 2-tailed unpaired Student’s t -tests. * p < .05, ** p < .01, ns denotes no significant difference. Transcripts per million (TPM): this is the most used standardized unit in RNA sequencing (RNA-seq) data. It represents the proportion of a certain gene’s transcripts among all transcripts in a specific sample or tissue. After standardization, it enables comparability between different samples and genes.

Article Snippet: Antibodies against RUNX2, SP7, NFATC1, and CTSK were purchased from Abcam, while the anti-GAPDH antibody and anti-FSTL1 antibody were obtained from Proteintech.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Comparison, RNA Sequencing

Journal: JBMR Plus

Article Title: Follistatin-like protein 1 (FSTL1) modulates bone remodeling and attenuates bone loss in a mouse model of postmenopausal osteoporosis

doi: 10.1093/jbmrpl/ziag037

Figure Lengend Snippet: Fstl1 haploinsufficiency disrupts bone homeostasis and aggravates OVX-induced bone loss. (A) Three-dimensional micro-CT reconstructions of femora. (B) quantitative micro-CT results. (C) Representative H&E images of distal femoral trabeculae. (D) Quantitative analysis of trabecular area (Tb.Ar) from H&E sections. (E) Osteocalcin (OCN) immunohistochemistry. (F) Quantitative analysis of osteoblast number per bone perimeter (N.Ob/B.Pm, 1/mm) in the indicated experimental groups. (G) TRAP staining depicting osteoclast distribution (red arrows). (H) Quantitative analysis of osteoclast number per bone perimeter (N.Oc/B.Pm, 1/mm) in the indicated experimental groups. Sample sizes: WT-sham ( n = 6), Fstl1 +/− sham ( n = 6), WT-OVX ( n = 6), and Fstl1 +/− OVX ( n = 6). Data are expressed as mean ± SD; group comparisons were conducted with 2-way ANOVA. * p < .05, ** p < .01, *** p < .001, **** p < .0001; ns, not significant.

Article Snippet: Antibodies against RUNX2, SP7, NFATC1, and CTSK were purchased from Abcam, while the anti-GAPDH antibody and anti-FSTL1 antibody were obtained from Proteintech.

Techniques: Micro-CT, Immunohistochemistry, Staining

Journal: JBMR Plus

Article Title: Follistatin-like protein 1 (FSTL1) modulates bone remodeling and attenuates bone loss in a mouse model of postmenopausal osteoporosis

doi: 10.1093/jbmrpl/ziag037

Figure Lengend Snippet: Fstl1 enhances the osteogenic differentiation potential of MC3T3-E1 cells. (A) Representative alkaline-phosphatase (ALP) staining at 7 d. (B) Quantitative ALP activity. (C) Representative Alizarin Red S (ARS) staining at 14 d. (D) Quantification of ARS absorbance (OD 562 ). (E-H) Relative mRNA expression of Runx2, Sp7, Ocn, and Opn at 14 d determined by RT-qPCR. (I) Western-blot bands for RUNX2 and SP7. (J and K) Densitometric ratios of RUNX2/GAPDH and SP7/GAPDH. Sample sizes: con ( n = 3), OE-Fstl1 ( n = 3), and sh-Fstl1 ( n = 3). Data are presented as mean ± SD; inter-group comparisons were performed using 1-way ANOVA. * p < .05, ** p < .01, *** p < .001, **** p < .0001; ns, not significant.

Article Snippet: Antibodies against RUNX2, SP7, NFATC1, and CTSK were purchased from Abcam, while the anti-GAPDH antibody and anti-FSTL1 antibody were obtained from Proteintech.

Techniques: Staining, Activity Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: JBMR Plus

Article Title: Follistatin-like protein 1 (FSTL1) modulates bone remodeling and attenuates bone loss in a mouse model of postmenopausal osteoporosis

doi: 10.1093/jbmrpl/ziag037

Figure Lengend Snippet: Fstl1 suppresses the osteoclast-differentiation activity of BMMs. (A) TRAP staining showing TRAP + multinucleated osteoclast morphology in each group. (B) Quantitative analysis of TRAP + osteoclast numbers. (C) Dual staining of F-actin rings (green) and DAPI (blue) illustrating the resorptive synapse structure of mature osteoclasts. (D) Average number of nuclei per TRAP + osteoclast. (E and F) Representative images of resorption pits on an osteoassay surface from each group and quantification of the total resorbed area per well (percentage of surface area). (G-J) Relative mRNA expression of Ctsk, Nfatc1, c-Fos, and Trap determined by RT-qPCR. (K) Western-blot bands for CTSK and NFATc1. (L and M) Densitometric ratios of NFATc1/GAPDH and CTSK/GAPDH. Sample sizes: con ( n = 3), OE-Fstl1 ( n = 3), and sh-Fstl1 ( n = 3). Data are presented as mean ± SD; inter-group comparisons were performed with 1-way ANOVA. * p < .05, ** p < .01, *** p < .001, **** p < .0001; ns, not significant.

Article Snippet: Antibodies against RUNX2, SP7, NFATC1, and CTSK were purchased from Abcam, while the anti-GAPDH antibody and anti-FSTL1 antibody were obtained from Proteintech.

Techniques: Activity Assay, Staining, Expressing, Quantitative RT-PCR, Western Blot

Journal: JBMR Plus

Article Title: Follistatin-like protein 1 (FSTL1) modulates bone remodeling and attenuates bone loss in a mouse model of postmenopausal osteoporosis

doi: 10.1093/jbmrpl/ziag037

Figure Lengend Snippet: Intraperitoneal administration of FSTL1 markedly restores bone mass in OVX mice. (A) Micro-CT three-dimensional reconstructions showing trabecular morphology in femoral cancellous bone. (B) Quantitative micro-CT parameters (BMD, BV/TV, Tb.Th, Tb.N, and Tb.Sp). (C) H&E staining illustrating trabecular architecture. (D) Quantitative analysis of trabecular area fraction (Tb.Ar%). (E) OCN immunohistochemistry depicting osteoblast distribution (red arrows). (F) Quantitative analysis of osteoblast number per bone perimeter (N.Ob/B.Pm, 1/mm) in the indicated experimental groups. (G) TRAP staining depicting osteoclast distribution (red arrows). (H) Quantitative analysis of osteoclast number per bone perimeter (N.Oc/B.Pm, 1/mm) in the indicated experimental groups. Sample sizes: OVX+PBS ( n = 6) and OVX+FSTL1 ( n = 6). Data are expressed as mean ± SD; 2-tailed unpaired Student’s t -tests were used for between-group comparisons. * p < .05, ** p < .01, *** p < .001; ns, not significant.

Article Snippet: Antibodies against RUNX2, SP7, NFATC1, and CTSK were purchased from Abcam, while the anti-GAPDH antibody and anti-FSTL1 antibody were obtained from Proteintech.

Techniques: Micro-CT, Staining, Immunohistochemistry