Journal: FEBS Open Bio

Article Title: Modeling atrial fibrosis in vitro —Generation and characterization of a novel human atrial fibroblast cell line

doi: 10.1002/2211-5463.12896

Figure Lengend Snippet: Morphological and immunocytochemical fibroblast identification. Representative brightfield and immunofluorescence images of the fibroblast markers vimentin, DDR2, collagen 1, and αSMA. The nuclei were stained with DAPI (blue). Upper panel) HVFs. Mid panel) HAFs. Lower panel) PAFs. The scale bars equal 50 µm.

Article Snippet: Using lentiviral transfer of Upcyte ® proliferation genes, a novel human atrial fibroblast cell line (HAF‐SRK01) was generated.

Techniques: Immunofluorescence, Staining

Journal: FEBS Open Bio

Article Title: Modeling atrial fibrosis in vitro —Generation and characterization of a novel human atrial fibroblast cell line

doi: 10.1002/2211-5463.12896

Figure Lengend Snippet: Induction of a cellular fibrosis phenotype in HAFs with TGF‐β. (A) Protein expression of phosphorylated SMAD2/3 after 72 h of TGF‐β stimulation (1, 3, 10 ng·mL −1 ) in HAFs ( n = 3 per concentration) and representative original WB below. (B) Protein expression of αSMA after 72 h of TGF‐β stimulation (1, 3, 10 ng·mL −1 ) in HAFs ( n = 3 per concentration) and representative original WB below. (C) Quantification of fibroblasts positive for fibrillary αSMA microfilaments after stimulation with 10 ng·mL −1 TGF‐β for 72 h and representative immunofluorescence staining of fibrillary αSMA upon TGF‐β stimulation. The scale bars equal 50 µm. (D) Soluble collagen secretion (left) and representative immunofluorescence image for deposited Collagen Iα1 (right) by HAFs upon stimulation with 10 ng·mL −1 TGF‐β ( n = 14 vs. 10). The scale bars equal 50 µm. (E) Proliferation curves of HAFs under control conditions ( n = 4) and upon stimulation with 10 ng·mL −1 TGF‐β ( n = 4). Cells were counted after 7 and 14 days. (F) 24‐h migration capacity of HAFs under control conditions ( n = 6) and upon stimulation with 10 ng·mL −1 TGF‐β ( n = 4). Data are presented as mean ± SEM. Differences between two groups were compared using Student’s t ‐test with Welch’s correction. Differences between multiple groups were compared with one‐way ANOVA with Newman–Keuls post‐test. * P < 0.05. ** P < 0.01. *** P < 0.001.

Article Snippet: Using lentiviral transfer of Upcyte ® proliferation genes, a novel human atrial fibroblast cell line (HAF‐SRK01) was generated.

Techniques: Expressing, Concentration Assay, Immunofluorescence, Staining, Control, Migration

Journal: FEBS Open Bio

Article Title: Modeling atrial fibrosis in vitro —Generation and characterization of a novel human atrial fibroblast cell line

doi: 10.1002/2211-5463.12896

Figure Lengend Snippet: Functional analysis of cardiac fibroblast subtypes. (A) Proliferation curves of HVFs ( n = 6), HAFs ( n = 7) and PAFs ( n = 21). Cells were cultured in DMEM (10% FCS, 1% penicillin–streptomycin) at 37 °C, 5% CO 2 , and counted after 7 and 14 days. (B) Quantification of immunostaining experiments for fibrillary αSMA protein abundance ( n HVF = 4, n HAF = 7, n PAF = 6). (C) Basal 24‐h migration capacity of HVFs ( n = 5), HAFs ( n = 5), and PAFs ( n = 6). Data are presented as mean ± SEM. Differences between multiple groups were compared with one‐way ANOVA with Newman–Keuls post‐test. * P < 0.05. ** P < 0.01. *** P < 0.001; n.s., not significant.

Article Snippet: Using lentiviral transfer of Upcyte ® proliferation genes, a novel human atrial fibroblast cell line (HAF‐SRK01) was generated.

Techniques: Functional Assay, Cell Culture, Immunostaining, Quantitative Proteomics, Migration

Journal: FEBS Open Bio

Article Title: Modeling atrial fibrosis in vitro —Generation and characterization of a novel human atrial fibroblast cell line

doi: 10.1002/2211-5463.12896

Figure Lengend Snippet: Adaption of HAF and PAF stiffness in response to different stiffness of the growth matrix. (A) HAFs present a typical fibroblast morphology when grown on CyPhyGels. Nuclei were stained with Hoechst (blue), F‐actin was stained with Phalloidin (red), and ɑSMA was stained in green. The scale bar equals 20 µm. (B) Representative force/ indentation curves used to calculate the stiffness (E eff ) of individual cells cultured on either stiff (black curve) or soft gels (grey curve). The force required to indent a cell on the stiff substrate is higher than on the soft substrate. (C) Measurements of HAF and PAF stiffness on soft (~2.7 kPa) and stiff (~4.6 kPa) CyPhyGels (36 ≤ n ≤ 57). Data are presented as mean ± SEM. Differences between multiple groups were compared with one‐way ANOVA with Newman–Keuls post‐test. *** P < 0.001.

Article Snippet: Using lentiviral transfer of Upcyte ® proliferation genes, a novel human atrial fibroblast cell line (HAF‐SRK01) was generated.

Techniques: Staining, Cell Culture

Journal: International Journal of Ophthalmology

Article Title: Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts

doi: 10.18240/ijo.2021.03.02

Figure Lengend Snippet: A: After exposure of HCFs to various concentrations of zymosan, the expression of Cx43 was examined by Western blot analysis; B: The immunoblots were subjected to densitometric analysis in order to determine the band intensity. The error bars represent the standard deviation. aP<0.05 (Dunnett's multiple comparison test) versus the control (no zymosan).

Article Snippet: Culture of Human Corneal Fibroblasts HCFs were purchased from ScienCell Research Laboratories (cat. no. #6520; Carlsbad, CA, USA) and cultured in MEM containing 10% FBS (5% CO 2 , 37°C).

Techniques: Expressing, Western Blot, Standard Deviation, Comparison, Control

Journal: International Journal of Ophthalmology

Article Title: Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts

doi: 10.18240/ijo.2021.03.02

Figure Lengend Snippet: A: After the exposure of HCFs to zymosan (600 µg/mL) for the indicated time, the expression of Cx43 was examined by Western blot analysis; B: The immunoblots were subjected to densitometric analysis in order to determine the band intensity. The error bars represent the standard deviation. aP<0.05 (Dunnett's multiple comparison test) versus the control (no zymosan).

Article Snippet: Culture of Human Corneal Fibroblasts HCFs were purchased from ScienCell Research Laboratories (cat. no. #6520; Carlsbad, CA, USA) and cultured in MEM containing 10% FBS (5% CO 2 , 37°C).

Techniques: Expressing, Western Blot, Standard Deviation, Comparison, Control

Journal: International Journal of Ophthalmology

Article Title: Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts

doi: 10.18240/ijo.2021.03.02

Figure Lengend Snippet: After HCFs were incubated with or without zymosan (600 µg/mL) for 24h, the Cx43 mRNA level in the cells was then determined by qRT-PCR analysis. The error bars represent the standard deviation. aP<0.05 (Student's t-test) versus the control (no zymosan).

Article Snippet: Culture of Human Corneal Fibroblasts HCFs were purchased from ScienCell Research Laboratories (cat. no. #6520; Carlsbad, CA, USA) and cultured in MEM containing 10% FBS (5% CO 2 , 37°C).

Techniques: Incubation, Quantitative RT-PCR, Standard Deviation, Control

Journal: International Journal of Ophthalmology

Article Title: Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts

doi: 10.18240/ijo.2021.03.02

Figure Lengend Snippet: A: After HCFs were treated with PD98059, SB203580, or JNK inhibitor II (at 1, 3, and 10 µmol/L), respectively, in the absence or presence of zymosan (600 µg/mL), the expression of Cx43 was then examined by Western blot analysis; B: Densitometric analysis was performed for the immunoblots to determine the band intensity. The error bars represent the standard deviation. aP<0.05 (Dunnett's test) versus the control (no zymosan); bP<0.05 (Dunnett's test) versus the corresponding control (zymosan only).

Article Snippet: Culture of Human Corneal Fibroblasts HCFs were purchased from ScienCell Research Laboratories (cat. no. #6520; Carlsbad, CA, USA) and cultured in MEM containing 10% FBS (5% CO 2 , 37°C).

Techniques: Expressing, Western Blot, Standard Deviation, Control

Journal: International Journal of Ophthalmology

Article Title: Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts

doi: 10.18240/ijo.2021.03.02

Figure Lengend Snippet: A: HCFs were treated with IKK2 inhibitor IV (1, 3, and 10 µmol/L) in the absence or presence of zymosan (600 µg/mL). The expression of Cx43 was then examined by Western blot analysis. B: The immunoblots were subjected to densitometric analysis in order to determine the band intensity. The error bars represent the standard deviation. aP<0.05 (Dunnett's test) versus the control (no zymosan). bP<0.05 (Dunnett's test) versus the corresponding control (zymosan only).

Article Snippet: Culture of Human Corneal Fibroblasts HCFs were purchased from ScienCell Research Laboratories (cat. no. #6520; Carlsbad, CA, USA) and cultured in MEM containing 10% FBS (5% CO 2 , 37°C).

Techniques: Expressing, Western Blot, Standard Deviation, Control

Journal: International Journal of Ophthalmology

Article Title: Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts

doi: 10.18240/ijo.2021.03.02

Figure Lengend Snippet: HCFs were treated with the ERK inhibitor PD98059, JNK inhibitor II, or IKK2 inhibitor IV (10 µmol/L), respectively, in the absence or presence of zymosan (600 µg/mL). A scrape-loading assay with Lucifer yellow was used to measure the GJIC activity. A: Representative fluorescence microscopy images of fixed cells from different treatments (scale bar, 100 µm); B: The maximum distances from the scrape to dye fluorescence were quantified to show the GJIC activity. The error bars represent the standard deviation. aP<0.05 (Dunnett's test) versus the control (no zymosan). bP<0.05 (Dunnett's test) versus the corresponding control (zymosan only).

Article Snippet: Culture of Human Corneal Fibroblasts HCFs were purchased from ScienCell Research Laboratories (cat. no. #6520; Carlsbad, CA, USA) and cultured in MEM containing 10% FBS (5% CO 2 , 37°C).

Techniques: Activity Assay, Fluorescence, Microscopy, Standard Deviation, Control