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11 week old wildtype mice were treated with either control IgG or neutralizing pan-TGFB antibody (1D11) (at 5mg/kg) three times over one week and were placed on a normal phosphate (0.6% Pi) or high phosphate (1.8% Pi) diet. a. Serum TGFB2 and b. serum iFGF23 were assessed by ELISA. c. Femoral Fgf23 , Tgfb1 , and Tgfb2 gene expression was assessed by RT-qPCR, with fold change relative to mice on a normal phosphate diet treated with IgG, normalized using expression of Rplp0. d. Kidney Cyp27b1 , Cyp24a1 , and Klotho gene expression was assessed by RT-qPCR, with fold change relative to mice on a normal phosphate diet treated with IgG, normalized using expression of Rplp0. e. Urine phosphate measured was assessed with colorimetric quantification. (n=3 for mice on 1.8% Pi diet treated with IgG due to loss of samples). f. Serum phosphate was assessed with colorimetric quantification. g. Serum <t>iPTH</t> was assessed by ELISA. h. Serum creatinine was assessed by ELISA. n= 4-6 biological replicates for all experiments. Significance was assessed by two-way ANOVA, and corrected for multiple comparisons using Šidák’s correction (*: adjusted p-value<0.05, ns: not significant).
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11 week old wildtype mice were treated with either control IgG or neutralizing pan-TGFB antibody (1D11) (at 5mg/kg) three times over one week and were placed on a normal phosphate (0.6% Pi) or high phosphate (1.8% Pi) diet. a. Serum TGFB2 and b. serum iFGF23 were assessed by ELISA. c. Femoral Fgf23 , Tgfb1 , and Tgfb2 gene expression was assessed by RT-qPCR, with fold change relative to mice on a normal phosphate diet treated with IgG, normalized using expression of Rplp0. d. Kidney Cyp27b1 , Cyp24a1 , and Klotho gene expression was assessed by RT-qPCR, with fold change relative to mice on a normal phosphate diet treated with IgG, normalized using expression of Rplp0. e. Urine phosphate measured was assessed with colorimetric quantification. (n=3 for mice on 1.8% Pi diet treated with IgG due to loss of samples). f. Serum phosphate was assessed with colorimetric quantification. g. Serum <t>iPTH</t> was assessed by ELISA. h. Serum creatinine was assessed by ELISA. n= 4-6 biological replicates for all experiments. Significance was assessed by two-way ANOVA, and corrected for multiple comparisons using Šidák’s correction (*: adjusted p-value<0.05, ns: not significant).
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11 week old wildtype mice were treated with either control IgG or neutralizing pan-TGFB antibody (1D11) (at 5mg/kg) three times over one week and were placed on a normal phosphate (0.6% Pi) or high phosphate (1.8% Pi) diet. a. Serum TGFB2 and b. serum iFGF23 were assessed by ELISA. c. Femoral Fgf23 , Tgfb1 , and Tgfb2 gene expression was assessed by RT-qPCR, with fold change relative to mice on a normal phosphate diet treated with IgG, normalized using expression of Rplp0. d. Kidney Cyp27b1 , Cyp24a1 , and Klotho gene expression was assessed by RT-qPCR, with fold change relative to mice on a normal phosphate diet treated with IgG, normalized using expression of Rplp0. e. Urine phosphate measured was assessed with colorimetric quantification. (n=3 for mice on 1.8% Pi diet treated with IgG due to loss of samples). f. Serum phosphate was assessed with colorimetric quantification. g. Serum iPTH was assessed by ELISA. h. Serum creatinine was assessed by ELISA. n= 4-6 biological replicates for all experiments. Significance was assessed by two-way ANOVA, and corrected for multiple comparisons using Šidák’s correction (*: adjusted p-value<0.05, ns: not significant).

Journal: bioRxiv

Article Title: TGFβ signaling regulates the response of the skeleton to phosphate

doi: 10.64898/2025.12.01.691659

Figure Lengend Snippet: 11 week old wildtype mice were treated with either control IgG or neutralizing pan-TGFB antibody (1D11) (at 5mg/kg) three times over one week and were placed on a normal phosphate (0.6% Pi) or high phosphate (1.8% Pi) diet. a. Serum TGFB2 and b. serum iFGF23 were assessed by ELISA. c. Femoral Fgf23 , Tgfb1 , and Tgfb2 gene expression was assessed by RT-qPCR, with fold change relative to mice on a normal phosphate diet treated with IgG, normalized using expression of Rplp0. d. Kidney Cyp27b1 , Cyp24a1 , and Klotho gene expression was assessed by RT-qPCR, with fold change relative to mice on a normal phosphate diet treated with IgG, normalized using expression of Rplp0. e. Urine phosphate measured was assessed with colorimetric quantification. (n=3 for mice on 1.8% Pi diet treated with IgG due to loss of samples). f. Serum phosphate was assessed with colorimetric quantification. g. Serum iPTH was assessed by ELISA. h. Serum creatinine was assessed by ELISA. n= 4-6 biological replicates for all experiments. Significance was assessed by two-way ANOVA, and corrected for multiple comparisons using Šidák’s correction (*: adjusted p-value<0.05, ns: not significant).

Article Snippet: Additional ELISAs were used to quantify serum TGFβ2 (DY7346-05, R&D Systems), serum intact parathyroid hormone (iPTH) (60-2305, QuidelOrtho), and urine creatinine concentrations (0430, Stanbio Laboratory).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Gene Expression, Quantitative RT-PCR, Expressing