rat primary cardiac fibroblasts  (STEMCELL Technologies Inc)

Journal: Journal of Thoracic Disease

Article Title: TIMP1 regulation of cardiac fibroblast proliferation via the TGF-β/Smad pathway in an in vitro model of atrial fibrillation

doi: 10.21037/jtd-2025-1088

Figure Lengend Snippet: Effect of ang II on the expression of TIMP1 , TGF-β/Smad pathway-related proteins, and α-SMA. (A) qRT-PCR was performed to assess the mRNA expression levels of TIMP1 in rat cardiac fibroblasts treated with different concentrations of ang II (1 nM, 10 nM, 100 nM, and 1 μM). (B) qRT-PCR was also performed to assess the mRNA expression levels of TIMP1 in rat atrial fibroblasts subjected to different durations of 1 μM of ang II treatment (12, 24, and 48 hours). (C) WB was used to detect the expression of TGF-β1, p-Smad2, p-Smad3, Smad2, Smad3 and α-SMA after 48 hours of treatment with 1 μM of ang II. (D) The results of WB analysis for the same proteins are shown. Each experiment was performed with at least three biological replicates. *, P<0.05. TIMP1 , tissue inhibitor of metalloproteinases-1; ang II, angiotensin II; mRNA, messenger RNA; qRT-PCR, quantitative real-time polymerase chain reaction; WB, Western blotting.

Article Snippet: Rat primary cardiac fibroblasts were purchased from Stemcell Biotechnology Co., Ltd. (cat. no. STM-CE-3303; Shanghai, China; https://www.stemrecell.com/primary-cell-rat-fibroblast/cardiacmuscle.html ).

Techniques: Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Western Blot

Journal: Journal of Thoracic Disease

Article Title: TIMP1 regulation of cardiac fibroblast proliferation via the TGF-β/Smad pathway in an in vitro model of atrial fibrillation

doi: 10.21037/jtd-2025-1088

Figure Lengend Snippet: TIMP1 modulation altered the ang II-mediated effects on rat atrial fibroblast viability and proliferation. (A,B) qRT-PCR and WB were used to detect the TIMP1 knockdown efficiency in rat cardiac fibroblasts treated with 1 μM of ang I for 48 hours. (C,D) Evaluation of TIMP1 overexpression efficiency in rat atrial fibroblasts after 48 hours of 1-μM ang II treatment. (E) qRT-PCR was used to detect the expression of TIMP1 in cardiac fibroblasts treated with 1 μM of ang II for 48 hours following knockdown or overexpression of TIMP1 . (F) Cell viability of rat cardiac fibroblasts was assessed by CCK-8 assay after treatment with 1 μM of ang II, either alone or in combination with negative control (NC), TIMP1 knockdown (si- TIMP1 -1), or TIMP1 overexpression (over- TIMP1 ). (G) Colony formation assay results of rat cardiac fibroblasts treated with control, 1 μM of ang II alone, ang II combined with NC, si- TIMP1 -1, or over- TIMP1 . Observed using a stereomicroscope and photographed with a scanner. Each experiment was performed with at least three biological replicates. *, P<0.05. TIMP1 , tissue inhibitor of metalloproteinases-1; ang II, angiotensin II; CCK-8, Cell Counting Kit-8; qRT-PCR, quantitative real-time polymerase chain reaction; WB, Western blotting.

Article Snippet: Rat primary cardiac fibroblasts were purchased from Stemcell Biotechnology Co., Ltd. (cat. no. STM-CE-3303; Shanghai, China; https://www.stemrecell.com/primary-cell-rat-fibroblast/cardiacmuscle.html ).

Techniques: Quantitative RT-PCR, Knockdown, Over Expression, Expressing, CCK-8 Assay, Negative Control, Colony Assay, Control, Cell Counting, Real-time Polymerase Chain Reaction, Western Blot

Journal: Journal of Thoracic Disease

Article Title: TIMP1 regulation of cardiac fibroblast proliferation via the TGF-β/Smad pathway in an in vitro model of atrial fibrillation

doi: 10.21037/jtd-2025-1088

Figure Lengend Snippet: Effects of TIMP1 regulation on fibrosis marker expression and TGF-β/Smad signaling in ang II-induced rat cardiac fibroblasts. (A-C) The mRNA expression levels of fibrotic markers, collagen I , collagen III , and α-SMA in rat cardiac fibroblasts were analyzed via qRT-PCR and WB assay. The experimental groups of rat cardiac fibroblasts were as follows: control, 1 μM of ang II, 1 μM of ang II + NC, 1 μM of ang II + si- TIMP1- 1, and 1 μM of ang II + over- TIMP1 . (D,E) WB analysis of the protein expression levels of the TGF-β/Smad3 signaling pathway components in rat cardiac fibroblasts, including TGF-β1, phosphorylated Smad2 (p-Smad2), phosphorylated Smad3 (p-Smad3), Smad2, and Smad3. The experimental groups of rat cardiac fibroblasts were as follows: control, 1 μM of ang II, 1 μM of ang II + NC, 1 μM of ang II + si- TIMP1- 1, and 1 μM of ang II + over- TIMP1 . Each experiment was performed with at least three biological replicates. *, P<0.05. TIMP1 , tissue inhibitor of metalloproteinases-1; ang II, angiotensin II; NC, negative control; qRT-PCR, quantitative real-time polymerase chain reaction; WB, Western blotting.

Article Snippet: Rat primary cardiac fibroblasts were purchased from Stemcell Biotechnology Co., Ltd. (cat. no. STM-CE-3303; Shanghai, China; https://www.stemrecell.com/primary-cell-rat-fibroblast/cardiacmuscle.html ).

Techniques: Marker, Expressing, Quantitative RT-PCR, Control, Negative Control, Real-time Polymerase Chain Reaction, Western Blot

Journal: iScience

Article Title: Single-cell sequencing of immune cells from the coronary sinus reveals immune mechanisms of the progression of persistent atrial fibrillation

doi: 10.1016/j.isci.2024.110127

Figure Lengend Snippet: The redefinition and function of cluster#2 (A) T-SNE projection output of FGFBP2-expressing cells in T cells (a); T-SNE projection output of FGFBP2-expressing cells in cluster #7 and #11 of T cells from CBMCs (b); T-SNE projection output of cells co-expressing TRDC with FGFBP2 (c); T-SNE projection output of FGFBP2-expressing cells in cluster #8 of T cells from CBMCs (d); T-SNE projection output of cells co-expressing CD4 with FGFBP2 (e); T-SNE projection output of FGFBP2+TRDC-CD4 − cells (Ftc-T cells) of T cells from CBMCs (f); T-SNE projection output of cells of cluster #2 (g). (B) Validation of the abundance of Ftc-T cells from CBMCs of the PerAF subjects by flow cytometry analysis in the validation cohort, N = 10 vs. 10. (C) Validation of the abundance of Ftc-T cells from CBMCs of the PSVT subjects by flow cytometry analysis in the validation cohort, N = 10 vs. 10. (D) Validation of FTR from CBMCs of PerAF (right) and PSVT (left) subjects by flow cytometry analysis in the validation cohort. N = 10 vs. 10, two-tailed unpaired Student’s t tests, p < 0.05 indicating statistical significance, ∗∗∗∗ p < 0.001, ∗∗∗∼∗∗ p < 0.01, ∗ p < 0.05. (E) Validation of FCR from CBMCs of PerAF (right) and PSVT (left) subjects by flow cytometry analysis in the validation cohort. N = 10 vs. 10, two-tailed unpaired Student’s t tests, p < 0.05 indicating statistical significance, ∗∗∗∗ p < 0.001, ∗∗∗∼∗∗ p < 0.01, ∗ p < 0.05. (F) Light microscopy view of Ftc-T cells co-cultured with CFs. (G) Co-culture of Ftc-T cells with CFs can promote the proliferation of CFs. Representative images of EDU incorporation (green) reflecting cell proliferation (a) and quantification of EDU-positive cells (b). Data are means ± SEM ( n = 3); ∗∗∗∗ p < 0.001, ∗∗∗∼∗∗ p < 0.01, ∗ p < 0.05. PerAF, persistent atrial fibrillation; PSVT, paroxysmal supra-ventricular tachycardia; T-SNE, T-distributed stochastic neighbor embedding; CBMCs, coronary sinus blood mononuclear cells; FTR, Ftc-T cells to the total T cells ratio; FCR, Ftc-T cells to the CBMCs ratio; CFs, cardiac fibroblasts.

Article Snippet: Primary rat cardiac fibroblasts (CFs) were obtained from Procell (Wuhan, China).

Techniques: Expressing, Biomarker Discovery, Flow Cytometry, Two Tailed Test, Light Microscopy, Cell Culture, Co-Culture Assay

Journal: iScience

Article Title: Single-cell sequencing of immune cells from the coronary sinus reveals immune mechanisms of the progression of persistent atrial fibrillation

doi: 10.1016/j.isci.2024.110127

Figure Lengend Snippet: Effect of IFI27 on cardiac fibroblasts (A) Differential expression of IFI27 between PerAF and PSVT subjects. a: differential expression of IFI27 in CBMCs between PerAF and PSVT subjects; b, differential expression of IFI27 in T cells between PerAF and PSVT subjects; c, differential expression of IFI27 in Ftc-T cells between PerAF and PSVT subjects. N = 10 vs. 10, Ftc-T cells: FGFBP2+TRDC-CD4 − T cells, two-tailed unpaired Student’s t tests, p < 0.05 indicating statistical significance, ∗∗∗∗ p < 0.001, ∗∗∗∼∗∗ p < 0.01, ∗ p < 0.05. (B–D) Validation of the differential expression of IFI27 in Ftc-T cells from CBMCs between PerAF and PSVT subjects. (B) Flow cytometry analysis of the differential expression of IFI27 in Ftc-T cells from CBMCs between PerAF (red) and PSVT (blue) subjects; (C) High expression of IFI27 in Ftc-T cells were identified in CBMCs of PerAF (right) than PSVT (left) subjects; (D) Validation of the differential expression of IFI27 in Ftc-T cells from CBMCs between PerAF and PSVT subjects by qRT-PCR in the validation cohort. N = 10 vs. 10, Ftc-T cells: FGFBP2+TRDC-CD4 − T cells, two-tailed unpaired Student’s t tests, p < 0.05 indicating statistical significance, ∗∗∗∗ p < 0.001, ∗∗∗∼∗∗ p < 0.01, ∗ p < 0.05. (E) Western blot (a, b) and qrt PCR (c) results showed that IFI27 protein over-expression and knockdown can promote and inhibit the expression of col1 and a-SMA in CFs. Data are means ± SEM ( n = 3); ∗∗∗∗ p < 0.001, ∗∗∗∼∗∗ p < 0.01, ∗ p < 0.05. (F) IFI27 protein overexpression and knockdown could promote and inhibit the proliferation of CFs. Representative images of EDU incorporation (green) reflecting cell proliferation (a) and quantification of EDU-positive cells (b). Data are means ± SEM ( n = 3); ∗∗∗∗ p < 0.001, ∗∗∗∼∗∗ p < 0.01, ∗ p < 0.05.CFs: cardiac fibroblasts.

Article Snippet: Primary rat cardiac fibroblasts (CFs) were obtained from Procell (Wuhan, China).

Techniques: Quantitative Proteomics, Two Tailed Test, Biomarker Discovery, Flow Cytometry, Expressing, Quantitative RT-PCR, Western Blot, Over Expression, Knockdown

Journal: iScience

Article Title: Single-cell sequencing of immune cells from the coronary sinus reveals immune mechanisms of the progression of persistent atrial fibrillation

doi: 10.1016/j.isci.2024.110127

Figure Lengend Snippet:

Article Snippet: Primary rat cardiac fibroblasts (CFs) were obtained from Procell (Wuhan, China).

Techniques: Recombinant, Reverse Transcription, Protein Extraction, Plasmid Preparation, Sequencing, Software