recombinant mouse basic fgf (bfgf)  (GenScript corporation)

Journal: Journal of Cardiovascular Development and Disease

Article Title: FGFR4 Is Required for Concentric Growth of Cardiac Myocytes during Physiologic Cardiac Hypertrophy

doi: 10.3390/jcdd11100320

Figure Lengend Snippet: FGFR4 inhibition abrogates cardiac myocyte hypertrophy induced by serum from fed pythons. Neonatal rat ventricular myocytes (NRVMs) were treated with media containing either vehicle, FGF2 (25 ng/mL), FGF23 (25 ng/mL), or serum from fasted or fed snakes (diluted to 2% of the total media volume), without or in combination with a small molecule inhibitor of FGFR4 (iFGFR4; BLU9931, 10 ng/mL), for 48 h. Compared to vehicle-treated NRVMs (Ctrl) or to NRVMs treated with serum from fasted pythons, serum collected from pythons twelve hours post-feeding (Python 12HPF; #### p < 0.0001) and three days post-feeding (Python 3DPF; #### p < 0.0001) induced a significant increase in myocyte area. When co-treated with iFGFR4 (textured bars), both Python 12HPF (**** p < 0.0001) and Python 3DPF (*** p = 0.0003) did not induce a significant increase in NRVM area when compared to the same treatment without iFGFR4. Python 12HPF ( $$$ p = 0.0003)- and Python 3DPF ( $$ p = 0.0016)-treated NRVMs had significantly increased myocyte area when compared to Python Fasted serum treatments. Treatment with recombinant FGF23 significantly increased myocyte area compared to Ctrl ( #### p < 0.0001), fasted python serum treatments ( $$$$ p < 0.0001), and FGF23 + iFGFR4 treatment (**** p < 0.0001) NRVMs. Treatment with recombinant FGF2 protein served as a positive control for hypertrophy ( ## p = 0.003). Treatment with serum from fasted and two-days-post-feeding water snakes (Water Snake Fasted, Water Snake 2DPF) served as negative controls Each individual color corresponds with the treatment indicated below the bars. # = versus Ctrl; * = versus same treatment + iFGFR4; $ = versus Python Fasted; 150 cells per condition; n = 3–4 independent isolations of NRVMs. All values are shown as mean ± SD.

Article Snippet: We used recombinant murine FGF2 (3139-FB) and FGF23 (2629-FG/CF) proteins from R&D Systems.

Techniques: Inhibition, Recombinant, Positive Control

Journal: Journal of Cardiovascular Development and Disease

Article Title: FGFR4 Is Required for Concentric Growth of Cardiac Myocytes during Physiologic Cardiac Hypertrophy

doi: 10.3390/jcdd11100320

Figure Lengend Snippet: Mouse primer sequences. The following oligonucleotides shown in 5′ to 3′ orientation were used as primers in quantitative real-time PCR analyses.

Article Snippet: We used recombinant murine FGF2 (3139-FB) and FGF23 (2629-FG/CF) proteins from R&D Systems.

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: Journal of Cardiovascular Development and Disease

Article Title: FGFR4 Is Required for Concentric Growth of Cardiac Myocytes during Physiologic Cardiac Hypertrophy

doi: 10.3390/jcdd11100320

Figure Lengend Snippet: Serum FGF23 levels are elevated in mice during late pregnancy. Wild-type and global FGFR4 knockout (FGFR4 −/− ) mice were investigated during late pregnancy (LP; 18 days post-mating) and compared to non-pregnant virgin littermates (NP). Serum FGF23 concentrations were measured via ELISA. Both ( A ) total FGF23 and ( B ) intact FGF23 were significantly elevated in the serum of wild-type (total **** p < 0.0001; intact ** p = 0.0014) and FGFR4 −/− mice (total **** p < 0.0001; intact **** p < 0.0001) when compared to their respective NP controls (n = 8–12 mice per genotype and timepoint). FGFR4 −/− LP mice exhibit significantly higher intact FGF23 compared to wild-type LP mice (** p = 0.0055). ( C ) The ratios of intact FGF23 over total FGF23 show no significant differences between groups (n = 8–12 mice per genotype and timepoint). ( D ) Cardiac tissue from LP mice showed significant decreases in Klotho expression in wild-type mice (* p = 0.0398) compared to wild-type NP mice, but there were no significant differences between NP and LP in FGFR4 −/− mice (n = 4–6 mice per genotype and timepoint). Cardiac tissue from wild-type mice in LP showed a significant decline in Fgfr1 mRNA levels (** p = 0.0026), while FGFR4 −/− mice had no significant change in expression when compared to their respective NP control mice (n = 6–9 mice per genotype and timepoint). Fgfr4 expression in cardiac tissue from wild-type mice in LP remained unaltered, compared to wild-type mice in LP (n = 7–8 mice per genotype and timepoint). RTqPCR analysis of tissue was conducted using Gapdh as a housekeeping gene. All values are shown as ±SD.

Article Snippet: We used recombinant murine FGF2 (3139-FB) and FGF23 (2629-FG/CF) proteins from R&D Systems.

Techniques: Knock-Out, Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: Journal of Cardiovascular Development and Disease

Article Title: FGFR4 Is Required for Concentric Growth of Cardiac Myocytes during Physiologic Cardiac Hypertrophy

doi: 10.3390/jcdd11100320

Figure Lengend Snippet: FGF23 is expressed in the heart during late pregnancy in mice. ( A ) In wild-type mice during late pregnancy (LP; 18 days post-mating), Fgf23 mRNA levels were not altered in bone or liver tissue compared to non-pregnancy (NP) (n = 5–6 mice per timepoint). ( B ) Cardiac tissue from LP mice showed significant increases in Fgf23 expression in both wild-type (* p = 0.0213) and FGFR4 knockout (FGFR4 −/− ) (* p = 0.0246) mice when compared to respective NP controls (n = 4–8 mice per genotype and timepoint). ( C ) In cardiac tissue of wild-type mice, no differences in the mRNA levels of the FGF23 regulatory genes Galnt3 , Furin , or Fam20c were observed between NP and LP (n = 7–8 mice per timepoint). ( D ) Bone tissue from wild-type mice showed no differences in mRNA levels of Galnt3 , Furin , or Fam20c (n = 5–6 mice per timepoint) in NP versus LP. RTqPCR analysis of tissue was conducted using Gapdh as a housekeeping gene. All values are shown as mean ± SD.

Article Snippet: We used recombinant murine FGF2 (3139-FB) and FGF23 (2629-FG/CF) proteins from R&D Systems.

Techniques: Expressing, Knock-Out

Journal: Journal of Cardiovascular Development and Disease

Article Title: FGFR4 Is Required for Concentric Growth of Cardiac Myocytes during Physiologic Cardiac Hypertrophy

doi: 10.3390/jcdd11100320

Figure Lengend Snippet: Pregnant mice have increased serum phosphorus and calcium despite FGF23 elevations. ( A ) Serum phosphorus was significantly elevated during late pregnancy (LP; 18 days post-mating) in wild-type mice (* p = 0.0196; n = 14–19 mice per timepoint) and FGFR4 knockout (FGFR4 −/− ) mice (**** p < 0.0001; n = 8–11) when compared to non-pregnant (NP) mice. FGFR4 −/− LP mice have significantly higher serum phosphorus compared to wild-type LP mice (** p = 0.0093; n = 8–14). ( B ) Serum calcium was also significantly elevated in pregnant wild-type (*** p = 0.0001; n = 14–19) and FGFR4 −/− mice (** p = 0.0026; n = 9–12), compared to NP controls. FGFR4 −/− LP mice have significantly higher serum calcium compared to wild-type LP mice (* p = 0.0364; n = 9–14). Renal tissue from wild-type mice in LP exhibited no alterations in the mRNA levels of ( C ) sodium-dependent phosphate transporters Slc34a1 and Slc34a3 or of ( D ) Klotho , Fgfr1 , or Fgfr4 (n = 8 mice per timepoint). RTqPCR analysis of tissue was conducted using Gapdh as a housekeeping gene. All values are shown as mean ± SD.

Article Snippet: We used recombinant murine FGF2 (3139-FB) and FGF23 (2629-FG/CF) proteins from R&D Systems.

Techniques: Knock-Out

Journal: Journal of Cardiovascular Development and Disease

Article Title: FGFR4 Is Required for Concentric Growth of Cardiac Myocytes during Physiologic Cardiac Hypertrophy

doi: 10.3390/jcdd11100320

Figure Lengend Snippet: FGFR4 inhibition abrogates cardiac myocyte hypertrophy induced by serum from fed pythons. Neonatal rat ventricular myocytes (NRVMs) were treated with media containing either vehicle, FGF2 (25 ng/mL), FGF23 (25 ng/mL), or serum from fasted or fed snakes (diluted to 2% of the total media volume), without or in combination with a small molecule inhibitor of FGFR4 (iFGFR4; BLU9931, 10 ng/mL), for 48 h. Compared to vehicle-treated NRVMs (Ctrl) or to NRVMs treated with serum from fasted pythons, serum collected from pythons twelve hours post-feeding (Python 12HPF; #### p < 0.0001) and three days post-feeding (Python 3DPF; #### p < 0.0001) induced a significant increase in myocyte area. When co-treated with iFGFR4 (textured bars), both Python 12HPF (**** p < 0.0001) and Python 3DPF (*** p = 0.0003) did not induce a significant increase in NRVM area when compared to the same treatment without iFGFR4. Python 12HPF ( $$$ p = 0.0003)- and Python 3DPF ( $$ p = 0.0016)-treated NRVMs had significantly increased myocyte area when compared to Python Fasted serum treatments. Treatment with recombinant FGF23 significantly increased myocyte area compared to Ctrl ( #### p < 0.0001), fasted python serum treatments ( $$$$ p < 0.0001), and FGF23 + iFGFR4 treatment (**** p < 0.0001) NRVMs. Treatment with recombinant FGF2 protein served as a positive control for hypertrophy ( ## p = 0.003). Treatment with serum from fasted and two-days-post-feeding water snakes (Water Snake Fasted, Water Snake 2DPF) served as negative controls Each individual color corresponds with the treatment indicated below the bars. # = versus Ctrl; * = versus same treatment + iFGFR4; $ = versus Python Fasted; 150 cells per condition; n = 3–4 independent isolations of NRVMs. All values are shown as mean ± SD.

Article Snippet: We used recombinant murine FGF2 (3139-FB) and FGF23 (2629-FG/CF) proteins from R&D Systems.

Techniques: Inhibition, Recombinant, Positive Control

Journal: bioRxiv

Article Title: KLF4 in smooth muscle cell-derived progenitor cells is essential for angiotensin II-induced cardiac inflammation and fibrosis

doi: 10.1101/2024.06.04.597485

Figure Lengend Snippet: Gli1 -Cre ERT -YFP mice were subjected to Angiotensin II (AngII)-induced cardiac hypertrophy and fibrosis, as described in the Methods section. (A) Masson’s trichrome staining of 4-week AngII or saline treated cardiac tissue; ECM/collagen deposition shown in blue. Representative 20x images of N=4 in each group are shown. (B) Cardiac tissue sections were immunofluorescently stained with anti-GFP antibody and subjected to label-free SHG imaging to visualize the collagen deposition (Red). YFP + cells were imaged (Green) and overlayed to examine their association with ECM/collagen deposition. Representative 40x images of saline (N=10) and AngII-treated (N=12) tissues are shown. ( C ) Cardiac tissue sections were immunofluorescently stained with anti-GFP (Green), anti-αSMA (Red) and DAPI. Representative 60x images of N=6 in each group are shown. (D-I) Cardiac tissue from 2-week AngII- or saline-treated mice were harvested and prepared for scRNA-seq, as described in the Methods section. (D) Uniform Manifold Approximation and Projection (UMAP) visualization is shown. Blue circle highlights the major cluster that is annotated as AdvSca1-SM and fibroblast (SM-Fib) clusters. (E) scRNA-seq data UMAP plot colored by treatment (Saline - blue; AngII - orange). Red circle highlights the AngII-induced shift. (F) YFP transcript positive cells were highlighted in green in the UMAP plot. (G&H) Composition of Saline and AngII-treated samples were visualized for all cells (G) and YFP + cells (H) of the scRNA-seq data. (I) Pathway analysis for genes up-regulated (top) and down-regulated (bottom) by AngII treatment in YFP + AdvSca1-SM cells.

Article Snippet: Sorted cells were plated in gelatin-coated plates with AdvSca1-SM media (α MEM [Gibco, Cat# 32571036]), 10% MSC qualified fetal bovine serum (FBS)(Thermofisher Cat #12662029), 1x Penicillin Streptomycin, 1ng/mL murine basic fibroblast growth factor (R&D systems 3139-FB), and 5ng/mL murine epidermal growth factor (R&D systems 2028-EG) at the density of 20,000/cm 2 .

Techniques: Staining, Saline, Imaging