plasma fgf21 concentrations  (R&D Systems)

Journal: Biochemical and biophysical research communications

Article Title: Growth hormone resistance induced by amino acid deprivation in fao cells is independent of FGF21.

doi: 10.1016/j.bbrc.2024.149811

Figure Lengend Snippet: Fig. 3. Effect of FGH21 addition on GH signaling. A, B) Immunoblotting of JAK2, p-JAK2, STAT5b, and HSP90. Fao cells were cultured in the Full or Zero medium with 0–10 [5] pM of recombinant FGF21 for 24 h. Cells were then treated with 100 nM GH or vehicle for 10 min. C) Igf1 mRNA levels were quantified using real-time qPCR. Values were normalized against the Actb mRNA level. Fao cells were cultured in the full medium with or without 100 nM GH and 0–105 pM of FGF21 for 24 h. Total RNA was isolated. Bar: means ± SEM (n = 3), *p < 0.05. (D) The expression of α-klotho (Kl), β-klotho (Klb), and FGF Receptor (Fgfr) 1–4 in Fao cells, rat primary hepatocytes, and rat tissue samples were analyzed using reverse transcription-PCR. (E) Immnoblotting of Erk and p-Erk. Fao cells were cultured in the full medium for 24 h. The cells were then treated with 0–104 pM of FGF21 or vehicle for 10 min.

Article Snippet: Plasma FGF21 concentrations were measured using a mouse/rat FGF21 ELISA kit (Proteintech, IL, USA).

Techniques: Western Blot, Cell Culture, Recombinant, Isolation, Expressing, Reverse Transcription

Journal: Experimental gerontology

Article Title: Impact of aging and caloric restriction on fibroblast growth factor 21 signaling in rat white adipose tissue.

doi: 10.1016/j.exger.2019.01.001

Figure Lengend Snippet: Fig. 1. Effect of aging and calorie restriction (CR) on plasma FGF21 concentration and Fgf21 expression in epididymal white adipose tissue (eWAT), liver, and soleus muscle (SOL) of Wistar rats. Total RNA was extracted from eWAT, liver, and SOL of AL-fed rats at 3.5 months of age. The mRNA expression of Fgf21 was quantified using RT-qPCR (n = 5–6). Tbp was used as a reference gene and to normalize the expression of the target gene (A). Total protein was extracted from eWAT, liver, and SOL of AL-fed rats at 3.5 months of age and quantified by western blotting, using anti-FGF21 and LMNB1 antibodies (B, upper panel). LMNB1 was used as a reference protein and to normalize target protein expression. The quantitative data (n = 3) are shown in B, lower panel. Plasma was collected from the four groups (AL-fed, AL-fasted, CR-fed and CR-fasted) of rats at 3.5, 9, and 24 months of age. FGF21 plasma concentration was measured by ELISA (n = 5–6) (C). Total RNA was extracted from the liver and eWAT from the four groups (AL-fed, AL-fasted, CR-fed and CR-fasted) of rats at 3.5, 9, and 24 months of age. The mRNA expression of Fgf21 in the liver (D) and eWAT (F) were quantified using RT-qPCR (n = 5–6), as described above (D, F). Total protein was extracted from the liver and eWAT of the four groups (AL-fed, AL-fast, CR- fed and CR-fast) of rats at 3.5, 9, and 24 months of age. FGF21 protein expression in the liver (E) and eWAT (G) was quantified by western blotting, as described above (E, G). The quantitative data (n = 5–6) are shown in the lower panels. Values shown in all panels are means ± SEM. *P < 0.05 vs. AL, †P < 0.05 vs. fed, analyzed using the Steel–Dwass test in the same age group. §P < 0.05, §§P < 0.01 vs. 3.5 months of age, analyzed using the Steel test within the AL-fed group. N.D.: not-detected.

Article Snippet: Plasma FGF21 concentration was measured using a Mouse/Rat FGF21 Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA), according to the manufacturer's protocol.

Techniques: Clinical Proteomics, Concentration Assay, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Experimental gerontology

Article Title: Impact of aging and caloric restriction on fibroblast growth factor 21 signaling in rat white adipose tissue.

doi: 10.1016/j.exger.2019.01.001

Figure Lengend Snippet: Fig. 2. Effects of aging and calorie restriction (CR) on the expression of FGF21-regulating genes in the liver of Wistar rats. Total RNA was extracted from the liver of the four groups (AL-fed, AL-fasted, CR-fed and CR-fasted) of rats at 3.5, 9, and 24 months of age. The mRNA ex- pression of Pparα (A), Chrebp (B), Atf4 (C) and Srebp- 1c (D) was quantified using RT-qPCR (n = 5–6), with Tbp being used as a reference gene and to normalize target gene expression. Values shown in all panels are means ± SEM. *P < 0.05 vs. AL, †P < 0.05 vs. fed, analyzed using the Steel–Dwass test within the same age group. §P < 0.05, §§P < 0.01 vs. 3.5 months of age, analyzed using the Steel test within the AL-fed group.

Article Snippet: Plasma FGF21 concentration was measured using a Mouse/Rat FGF21 Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA), according to the manufacturer's protocol.

Techniques: Expressing, Quantitative RT-PCR, Targeted Gene Expression

Journal: Experimental gerontology

Article Title: Impact of aging and caloric restriction on fibroblast growth factor 21 signaling in rat white adipose tissue.

doi: 10.1016/j.exger.2019.01.001

Figure Lengend Snippet: Fig. 3. Effects of aging and calorie restriction (CR) on FGF21-regulating gene expression in the epidi- dymal white adipose tissue (eWAT) of Wistar rats. Total RNA was extracted from the eWAT of four groups (AL-fed, AL-fasted, CR-fed and CR-fasted) of rats at 3.5, 9, and 24 months of age. The mRNA ex- pression of Pparγ (A), Chrebp (B), Atf4 (C) and Srebp- 1c (D) was quantified using RT-qPCR (n = 5–6), with Tbp being used as a reference gene and to normalize target gene expression. Values shown in all panels are means ± SEM. *P < 0.05 vs. AL, †P < 0.05 vs. fed, analyzed using the Steel–Dwass test within the same age group. §P < 0.05, §§P < 0.01 vs. 3.5 months of age, analyzed using the Steel test within the AL-fed group.

Article Snippet: Plasma FGF21 concentration was measured using a Mouse/Rat FGF21 Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA), according to the manufacturer's protocol.

Techniques: Gene Expression, Quantitative RT-PCR, Targeted Gene Expression

Journal: Experimental gerontology

Article Title: Impact of aging and caloric restriction on fibroblast growth factor 21 signaling in rat white adipose tissue.

doi: 10.1016/j.exger.2019.01.001

Figure Lengend Snippet: Fig. 4. FGF21 signaling in rat epididymal white adipose tissue (eWAT) decreased with age, but caloric restriction (CR) ameliorated this effect. Total RNA was extracted from the eWAT of 3.5-month-old AL-fed, 9-month-old AL-fed and CR-fed, and 24-month-old AL-fed and CR-fed, groups of rats. The mRNA expression of Fgfr1 (A), Klb (B), Glut1 (C), Glut4 (D), and Pgc-1α (E) was quantified using RT-qPCR (n = 5–6), using Tbp as a reference gene and to normalize target gene expression (A–E). Total protein was extracted from the eWAT of 3.5-month-old AL-fed, 9-month-old AL-fed and CR-fed, and 24-month-old AL-fed and CR-fed, groups of rats. FGFR1, KLB, and GLUT1 protein expression was quantified using western blotting (F, left panel), with LMNB1 being used as a reference protein and to normalize target protein expression. The quantitative data (n = 5–6) are shown in the middle and right panels (F). Values shown in all panels are means ± SEM. *P < 0.05 vs. AL within the same age group, analyzed using the Mann–Whitney U test. §P < 0.05, §§P < 0.01 vs. 3.5-month-old rats, analyzed using the Steel test within the AL-fed group.

Article Snippet: Plasma FGF21 concentration was measured using a Mouse/Rat FGF21 Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA), according to the manufacturer's protocol.

Techniques: Expressing, Quantitative RT-PCR, Targeted Gene Expression, Western Blot, MANN-WHITNEY

Journal: Experimental gerontology

Article Title: Impact of aging and caloric restriction on fibroblast growth factor 21 signaling in rat white adipose tissue.

doi: 10.1016/j.exger.2019.01.001

Figure Lengend Snippet: Fig. 5. FGF21 overexpression altered expression of FGF21 downstream targets in 3T3-L1 adipocytes. Mock and FGF21 OE pre-adipocytes were differentiated into adipocytes and studies at day 12. Total RNA was extracted from the adipocytes and mRNA expression of Fgf21 (A), Fgfr1 (B), Klb (C), Glut1 (D), Glut4 (E), and Pgc-1α (F) were quantified using RT-qPCR (n = 4). Rps18 was used as a reference gene and to normalize target gene expression (A–F). Total cell lysates were analyzed by western blotting using anti-FGF21, FGFR1, KLB, GLUT1, and LMNB1 antibodies, with LMNB1 being used as a reference protein and to normalize target protein expression (G, left panel). The quantitative data (n = 4) are shown in the middle and right panels (G). Values shown in all panels are means ± SEM. *P < 0.05, analyzed using the Mann–Whitney U test.

Article Snippet: Plasma FGF21 concentration was measured using a Mouse/Rat FGF21 Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA), according to the manufacturer's protocol.

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Targeted Gene Expression, Western Blot, MANN-WHITNEY