recombinant mouse fibroblast growth factors fgf basic protein  (R&D Systems)

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

Journal: eLife

Article Title: Metabolic memory of Δ9-tetrahydrocannabinol exposure in pluripotent stem cells and primordial germ cells-like cells

doi: 10.7554/elife.88795

Figure Lengend Snippet: Figure 1. Δ9-THC exposure induces the proliferation of ESCs but not EpiLCs (A) Diagram illustrating Δ9-THC exposure scheme and experimental strategy. bFGF: basic fibroblast growth factor, ESCs, embryonic stem cells; EpiLCs, epiblast-like cells; LIF, leukemia inhibitory factor. (B, E, H) Whisker boxplot indicating the median cellular viability of stem cells exposed to the different Δ9-THC doses and associated errors. (C, F, I) Whisker boxplot indicating the median number of viable cells exposed to the different Δ9-THC doses indicated and associated errors. (D, G, J) Whisker boxplot indicating the median percentage of BrdU-stained cells exposed to the different Δ9-THC doses and associated errors. ESCs exposed cells are presented in (B, C and D). EpiLCs exposed cells deriving from unexposed ESCs are presented in (E, F and G). EpiLCs exposed cells deriving from exposed ESCs are presented in (H, I and J). At least three independent biological repeats with three technical replicates (N=3, n=3). Statistical significance: *(p<0.05), **(p<0.01), ***(p<0.001), ****(p<0.0001).

Article Snippet: Differentiation of ESCs to EpiLCs was performed by seeding the cells on Human Plasma Fibronectin (HPF)- coated plates [16.7 μg/mL; 33016015; Thermo Fisher] in the presence of EpiLC induction medium (N2B27 medium containing activin A [20 ng/mL; 50- 398- 465; Thermo Fisher]), basic fibroblast growth factor (bFGF) [12 ng/mL; 3139FB025; R&D Systems], and KnockOut Serum Replacement [KSR, 1%; Thermo Fisher].

Techniques: Whisker Assay, Staining