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

Journal: Poultry science

Article Title: MAPK signaling pathway participates in the regulation of intestinal phosphorus and calcium absorption in broiler chickens via 1,25-dihydroxyvitamin D 3 .

doi: 10.1016/j.psj.2024.104052

Figure Lengend Snippet: Figure 1. The mRNA and protein levels of jejunal NaPi-IIb and CaBP-D28k and the protein levels of p-ERK and p-p38MAPK in broiler chick- ens (9−14 days old) after intraperitoneal injection of saline (control) or 1,25(OH)2D3 (Experiment 2). (A) NaPi-IIb mRNA expression; (B) CaBP- D28k mRNA expression; (C) NaPi-IIb and CaBP-D28k protein expression; (E) p-ERK and p-p38MAPK protein expression. The values of the mRNA levels of NaPi-IIb and CaBP-D28k are the means of 4 replicates (2 broilers per replicate) (n = 4) § standard deviation (SD). The protein abundances of NaPi-IIb, CaBP-D28k, p-ERK, and p-p38MAPK are the means of 4 replicates (1 broiler per replicate) (n = 4) § SD. a-b Values with different superscripts are different (P < 0.05).

Article Snippet: The PVDF membrane was then blocked with 5% nonfat powdered milk in Tris-buffered saline supplemented with tween-20 (TBST) at 37°C for 1 h. Subsequently, the membranes were incubated overnight at 4°C with primary antibodies against GAPDH (1:10,000, 60004-1- Ig, Proteintech, Wuhan, China), NaPi-IIb (1:1,000, A9460, ABclonal, Wuhan, China), CaBP-D28k (1:5,000, 14479-1-AP, Proteintech, Wuhan, China), p-ERK (1:1,000, 9101S, Cell Signaling Technology, Boston, USA), t-ERK (1:8,000, 11257-1-AP-50, Proteintech, Wuhan, China), p-p38MAPK (1:1,000, 9216S, Cell Signaling Technology, Boston, USA), and t-p38MAPK (1:1,000, 14064-1-AP, Proteintech, Wuhan, China).

Techniques: Injection, Saline, Control, Expressing, Standard Deviation

Journal: Poultry science

Article Title: MAPK signaling pathway participates in the regulation of intestinal phosphorus and calcium absorption in broiler chickens via 1,25-dihydroxyvitamin D 3 .

doi: 10.1016/j.psj.2024.104052

Figure Lengend Snippet: Figure 2. The mRNA and protein abundances of jejunal NaPi-IIb and CaBP-D28k in broiler chickens (9−17 days old) after intraperitoneal injection of saline (control) or the ERK inhibitor PD98059 (Experiment 3). (A) NaPi-IIb mRNA expression; (B) CaBP-D28k mRNA expression; (C) NaPi-IIb and CaBP-D28k protein expression. The values of the NaPi-IIb and CaBP-D28k mRNA abundances are the means of 4 replicates (2 broilers per replicate) (n = 4) § SD. The values of the NaPi-IIb and CaBP-D28k protein abundances are the means of 4 cages (1 broiler per cage) (n = 4) § SD. a-b Values with different superscripts are different (P < 0.05).

Article Snippet: The PVDF membrane was then blocked with 5% nonfat powdered milk in Tris-buffered saline supplemented with tween-20 (TBST) at 37°C for 1 h. Subsequently, the membranes were incubated overnight at 4°C with primary antibodies against GAPDH (1:10,000, 60004-1- Ig, Proteintech, Wuhan, China), NaPi-IIb (1:1,000, A9460, ABclonal, Wuhan, China), CaBP-D28k (1:5,000, 14479-1-AP, Proteintech, Wuhan, China), p-ERK (1:1,000, 9101S, Cell Signaling Technology, Boston, USA), t-ERK (1:8,000, 11257-1-AP-50, Proteintech, Wuhan, China), p-p38MAPK (1:1,000, 9216S, Cell Signaling Technology, Boston, USA), and t-p38MAPK (1:1,000, 14064-1-AP, Proteintech, Wuhan, China).

Techniques: Injection, Saline, Control, Expressing

Journal: Poultry science

Article Title: MAPK signaling pathway participates in the regulation of intestinal phosphorus and calcium absorption in broiler chickens via 1,25-dihydroxyvitamin D 3 .

doi: 10.1016/j.psj.2024.104052

Figure Lengend Snippet: Figure 3. The mRNA and protein abundances of jejunal NaPi-IIb and CaBP-D28k in broiler chickens (9−17 days old) after intraperitoneal injection of saline (control) or the p38MAPK inhibitor SB203580 (Experiment 4). (A) NaPi-IIb mRNA expression; (B) CaBP-D28k mRNA expres- sion; (C) NaPi-IIb and CaBP-D28k protein expression. The values of NaPi-IIb and CaBP-D28k mRNA abundances are the means of 4 replicates (2 broilers per replicate) (n = 4) § SD. The values of NaPi-IIb and CaBP-D28k protein abundances are the means of 4 cages (1 broiler per cage) (n = 4) § SD. a-b Values with different superscripts are different (P < 0.05).

Article Snippet: The PVDF membrane was then blocked with 5% nonfat powdered milk in Tris-buffered saline supplemented with tween-20 (TBST) at 37°C for 1 h. Subsequently, the membranes were incubated overnight at 4°C with primary antibodies against GAPDH (1:10,000, 60004-1- Ig, Proteintech, Wuhan, China), NaPi-IIb (1:1,000, A9460, ABclonal, Wuhan, China), CaBP-D28k (1:5,000, 14479-1-AP, Proteintech, Wuhan, China), p-ERK (1:1,000, 9101S, Cell Signaling Technology, Boston, USA), t-ERK (1:8,000, 11257-1-AP-50, Proteintech, Wuhan, China), p-p38MAPK (1:1,000, 9216S, Cell Signaling Technology, Boston, USA), and t-p38MAPK (1:1,000, 14064-1-AP, Proteintech, Wuhan, China).

Techniques: Injection, Saline, Control, Expressing