Journal: Acta physiologica (Oxford, England)

Article Title: Short-term fasting of mice elevates circulating fibroblast growth factor 23 (FGF23).

doi: 10.1111/apha.14049

Figure Lengend Snippet: FIGURE 5 Altered NaPiIIa (Slc34a1), 24-hydroxylase (Cyp24a1) expression and αKlotho formation in fasted mice. Arithmetic means ± SEM (n = 5) of relative Slc34a1 (A), Cyp24a1 (C), 1α-hydroxylase (Cyp27b1) (D), and αKlotho (Kl) (E) mRNA abundance normalized to hypoxanthine guanine phosphoribosyl transferase (Hprt) expression in kidneys from fed mice and mice fasted for 16 h. Representative Western Blots and arithmetic means ± SEM of renal NaPiIIa (⁓69 kDa, isoform X1: ⁓59 kDa, isoform X2: ⁓48 kDa; B: n = 7) and Klotho (⁓130 to 135 kDa; F: n = 5) protein abundance relative to loading control β-tubulin (⁓55 kDa) in fed and fasted mice. For all panels, n denotes the number of specimens included. There were no dropouts in any analysis. *p < 0.05, †p < 0.01, ‡p < 0.001 indicate significant differences compared to fed mice. ns, non-significant. (A: Mann–Whitney U test, B–F: unpaired t-test).

Article Snippet: Proteins from UMR106 cells, NRVM, kidney, heart, thymus, bone marrow, pancreas (each 30 μg), or bone (10 μg) were subjected to standard Western Blot procedure using 10% or 12% SDS- PAGE gels or 4%– 20% precast SDS- PAGE gels (for FGF23 detection in NRVM), respectively, and the following primary antibodies: anti- NFκB- p65 (#8242, Cell Signaling Technology), anti- phospho- NFκB- p65 (#3033, Cell Signaling Technology), anti- FGF23 (MAB26291, R&D Systems), anti- FGF23 (#6320, Immutopics), anti- NaPiIIa (NBP2- 13328, Novus Biologicals, Bio- Techne), anti- Klotho (AF1819, R&D Systems), anti- β- actin ((#3700 (for lysates from UMR106 cells) and #8457 (for mouse tissue lysates), Cell Signaling Technology)), anti- GAPDH (#2118, Cell Signaling Technology), or anti- β- tubulin (#2146, Cell Signaling Technology).

Techniques: Expressing, Western Blot, Quantitative Proteomics, Control, MANN-WHITNEY

Journal: Acta physiologica (Oxford, England)

Article Title: Short-term fasting of mice elevates circulating fibroblast growth factor 23 (FGF23).

doi: 10.1111/apha.14049

Figure Lengend Snippet: FIGURE 5 Altered NaPiIIa (Slc34a1), 24-hydroxylase (Cyp24a1) expression and αKlotho formation in fasted mice. Arithmetic means ± SEM (n = 5) of relative Slc34a1 (A), Cyp24a1 (C), 1α-hydroxylase (Cyp27b1) (D), and αKlotho (Kl) (E) mRNA abundance normalized to hypoxanthine guanine phosphoribosyl transferase (Hprt) expression in kidneys from fed mice and mice fasted for 16 h. Representative Western Blots and arithmetic means ± SEM of renal NaPiIIa (⁓69 kDa, isoform X1: ⁓59 kDa, isoform X2: ⁓48 kDa; B: n = 7) and Klotho (⁓130 to 135 kDa; F: n = 5) protein abundance relative to loading control β-tubulin (⁓55 kDa) in fed and fasted mice. For all panels, n denotes the number of specimens included. There were no dropouts in any analysis. *p < 0.05, †p < 0.01, ‡p < 0.001 indicate significant differences compared to fed mice. ns, non-significant. (A: Mann–Whitney U test, B–F: unpaired t-test).

Article Snippet: Proteins from UMR106 cells, NRVM, kidney, heart, thymus, bone marrow, pancreas (each 30 μg), or bone (10 μg) were subjected to standard Western Blot procedure using 10% or 12% SDS- PAGE gels or 4%– 20% precast SDS- PAGE gels (for FGF23 detection in NRVM), respectively, and the following primary antibodies: anti- NFκB- p65 (#8242, Cell Signaling Technology), anti- phospho- NFκB- p65 (#3033, Cell Signaling Technology), anti- FGF23 (MAB26291, R&D Systems), anti- FGF23 (#6320, Immutopics), anti- NaPiIIa (NBP2- 13328, Novus Biologicals, Bio- Techne), anti- Klotho (AF1819, R&D Systems), anti- β- actin ((#3700 (for lysates from UMR106 cells) and #8457 (for mouse tissue lysates), Cell Signaling Technology)), anti- GAPDH (#2118, Cell Signaling Technology), or anti- β- tubulin (#2146, Cell Signaling Technology).

Techniques: Expressing, Western Blot, Quantitative Proteomics, Control, MANN-WHITNEY

Journal: Scientific Reports

Article Title: The intestinal phosphate transporter NaPi-IIb (Slc34a2) is required to protect bone during dietary phosphate restriction

doi: 10.1038/s41598-017-10390-2

Figure Lengend Snippet: Hormonal adaptation to dietary Pi is similar in both genotypes. Circulating levels of 1,25-(OH) 2 vitamin D 3 (A) , PTH (D) and intact FGF23 (E) were measured in plasma collected from wild type (WT) and NaPi-IIb −/− mice (KO) fed diets containing high (H) or low (L) amounts of Pi. The high Pi diet was provided for 3 days (3d) whereas the low Pi diet was provided for 3 (3d) and 14 days (14d). Renal mRNA levels of Cyp27b1 (B) and renal protein abundance of Cyp24a1 (C) were measured after 14 days of Pi restriction. Data is presented as mean + SEM (n = 10), and was analyzed by ANOVA-Bonferroni. Significant differences are indicated as: b p < 0.01 and c p < 0.001, were significances refer to high dietary Pi groups (no differences between genotypes were observed).

Article Snippet: Tris buffered saline (TBS) containing 5% fat free powder milk was used to block membranes for 30 minutes at room temperature prior incubation overnight at 4 °C with primary antibodies against NaPi-IIa , NaPi-IIb , NaPi-IIc , TRPV5 , CaSR (Thermofisher Scientific), Calbindin D28k (SWANT, Marly, Switzerland), Cyp24a1 (Protein Tech, Manchester, United Kingdom), and β-actin (Sigma-Aldrich, Buchs, Switzerland).

Techniques: Clinical Proteomics, Quantitative Proteomics