Journal: Cell

Article Title: Structural and Functional Analysis of the D614G SARS-CoV-2 Spike Protein Variant

doi: 10.1016/j.cell.2020.09.032

Figure Lengend Snippet:

Article Snippet: SMM-293 TII serum-free media , SinoBiological , M293TI.

Techniques: Binding Assay, Recombinant, Transfection, Luciferase, Bradford Protein Assay, Plasmid Preparation, Software, Single Particle, Microscopy

Journal: Circulation Research

Article Title: Regulation of Vascular Smooth Muscle Cell Turnover by Endothelial Cell–Secreted MicroRNA-126

doi: 10.1161/circresaha.113.280883

Figure Lengend Snippet: Figure 1. Atheroprotective shear stress (SS) to endothelial cells (ECs) suppresses microRNA-126 (miR-126) in the cocultured smooth muscle cells (SMCs). A, Schematic diagrams of EC/SMC coculture and flow system. B, Human umbilical vein ECs and human umbilical artery SMCs were seeded on the lower and upper sides of a membrane, respectively (EC/SMC). Controls had SMCs but no ECs on the other side (Ø/SMC). The temporal expression levels of primary (pri–miR-126) and mature (miR-126) forms of miR-126 were determined by quantitative reverse transcription–polymerase chain reaction. *P<0.05 vs Ø/SMC 0-hour static. C, The levels of miR-126 in the 24-hour static or sheared media. *P<0.05 vs static control. D, The levels of miR-126 in SMCs incubated with medium 199 supplemented with 2% fetal bovine serum (control [CL] media) or conditioned medium from static or sheared ECs (24-hour shearing) (EC-CM). *P<0.05 vs CL media. E, miR-126 expression in EC monoculture or SMC coculture for 24 hours. F, miR-126 expression in ECs exposed to laminar SS (LSS) for 24 hours. G, The levels of pri–miR-126 and miR-126 in EC coculture with SMCs under 24–hour static or LSS conditions.

Article Snippet: Human umbilical vein ECs (HUVECs) were cultured in medium 199 (Gibco) supplemented with 10% fetal bovine serum (FBS) (Omega Scientific) and 10% Endothelial Growth Medium (Cell Applications).

Techniques: Shear, Membrane, Expressing, Reverse Transcription, Polymerase Chain Reaction, Control, Incubation

Journal: Circulation Research

Article Title: Regulation of Vascular Smooth Muscle Cell Turnover by Endothelial Cell–Secreted MicroRNA-126

doi: 10.1161/circresaha.113.280883

Figure Lengend Snippet: Figure 2. Endothelial cells (ECs) and shear stress (SS) regulate expressions of microRNA-126 (miR-126) targets in smooth muscle cells (SMCs). A, The mRNA levels of forkhead box O3 (FOXO3), B-cell lymphoma 2 (BCL2), and insulin receptor substrate 1 (IRS1) in Argonaute2 (Ago2) immunocomplexes in SMCs 48 hours after transfection with microRNA-126 mimics (PRE126) or anti–microRNA-126 inhibitor (AM126). *P<0.05 vs cells transfected with negative controls (negative control mimics [PREC] or negative control inhibitor [AMC]). B, The relative luciferase levels in SMCs cotransfected with PRE126 (left), AM126 (right), or the negative control and luciferase reporter plasmids: empty vector only, wild-type (WT) 3′-untranslated region (3′UTR) of FOXO3, BCL2, or IRS1 and mutants of 3′UTR of FOXO3, BCL2, or IRS1. C, The relative luciferase levels in SMCs transfected with the indicated luciferase reporter plasmids and then incubated with control media or EC-conditioned media (CM). *P<0.05 vs cells transfected with empty vector. #P<0.05 vs cells transfected with plasmids harboring the WT 3′UTRs. D, The protein levels of target genes in SMCs monocultured (Ø/SMC) or cocultured with ECs (EC/SMC) kept under static condition or exposed to laminar shear stress (LSS). Images are representative of triplicate experiments with similar results.

Article Snippet: Human umbilical vein ECs (HUVECs) were cultured in medium 199 (Gibco) supplemented with 10% fetal bovine serum (FBS) (Omega Scientific) and 10% Endothelial Growth Medium (Cell Applications).

Techniques: Shear, Transfection, Negative Control, Luciferase, Plasmid Preparation, Incubation, Control

Journal: Circulation Research

Article Title: Regulation of Vascular Smooth Muscle Cell Turnover by Endothelial Cell–Secreted MicroRNA-126

doi: 10.1161/circresaha.113.280883

Figure Lengend Snippet: Figure 3. Endothelial cells (ECs) transfer exogenous microRNA-126 (miR-126) to smooth muscle cells (SMCs). SMCs were incubated with conditioned media (CM) derived from ECs transfected with biotinylated synthetic miR-126 (B-miR-126) or no-miR mock control (Mock) for 3 hours. A, The levels of B-miR-126 in ECs or the EC-CM were determined by pull-down experiments with streptavidin- agarose beads followed by quantitative reverse transcription–polymerase chain reaction, (B) and B-miR-126 in ECs and SMCs was detected with QDot-605–streptavidin conjugates. *P<0.05 vs mock control. C and D, SMCs were cocultured with ECs transfected with B-miR-126 or mock control for 24 hours; the levels of B-miR-126 in SMCs were determined by Northern blot (C) or pull-down experiments with streptavidin-agarose beads (D). *P<0.05 vs mock control.

Article Snippet: Human umbilical vein ECs (HUVECs) were cultured in medium 199 (Gibco) supplemented with 10% fetal bovine serum (FBS) (Omega Scientific) and 10% Endothelial Growth Medium (Cell Applications).

Techniques: Incubation, Derivative Assay, Transfection, Control, Reverse Transcription, Polymerase Chain Reaction, Northern Blot

Journal: Circulation Research

Article Title: Regulation of Vascular Smooth Muscle Cell Turnover by Endothelial Cell–Secreted MicroRNA-126

doi: 10.1161/circresaha.113.280883

Figure Lengend Snippet: Figure 4. Functional microRNA-126 (miR-126) transmission from endothelial cells (ECs) to smooth muscle cells (SMCs) is independent of vesicles. A, The levels of miR-126 in SMCs incubated with control (CL) media or the EC-conditioned media (CM) treated with DNase I, RNase, or proteinase K (PK) for 3 hours. *P<0.05 vs CL media. B, The levels of miR-126 in ECs transfected with control small interfering (si)RNA; siCL) or siRNA against Drosha (siDROSHA), Dicer (siDICER1) or Argonaute2 (siEIF2C2) or in SMCs incubated with CL media or EC-CM derived from those transfected ECs. C, The levels of miR-126 in SMCs incubated EC-CM derived from ECs transfected with anti–miR-126 inhibitor anti–miR-126 inhibitor (AM126), miR-126 mimics (PRE126), or the negative controls. *P<0.05 vs siCL or CL media. #P<0.05 vs the negative control inhibitor (AMC) or negative control mimics (PREC) EC-CM. D, The levels of miR-126 in SMCs cocultured with lung ECs isolated from wild-type (WT) or miR-126 knockout mice (KO). *P<0.05 vs Ø/SMC (SMC monocultured). EC-CM was ultracentrifuged to fractionate the components in spin-down pellets (Pellet; which was reconstituted with M199), or the remaining supernatant (Super). The levels of miR-126 in (E) the fractionated EC-CM or in (F) SMCs incubated with total or fractionated EC-CM for 3 hours. G, The transcript levels or (H), protein levels of indicated genes in SMCs incubated with the fractioned EC-CM. Images are representative of triplicates with similar results. *P<0.05 vs CL media.

Article Snippet: Human umbilical vein ECs (HUVECs) were cultured in medium 199 (Gibco) supplemented with 10% fetal bovine serum (FBS) (Omega Scientific) and 10% Endothelial Growth Medium (Cell Applications).

Techniques: Functional Assay, Transmission Assay, Incubation, Control, Transfection, Derivative Assay, Negative Control, Isolation, Knock-Out

Journal: Circulation Research

Article Title: Regulation of Vascular Smooth Muscle Cell Turnover by Endothelial Cell–Secreted MicroRNA-126

doi: 10.1161/circresaha.113.280883

Figure Lengend Snippet: Figure 5. Argonaute2 (Ago2) associates with microRNA-126 (miR-126) and facilitates its transmission. A, The levels of miR-126 in endothelial cell–conditioned media (EC-CM) immunocomplexes of Ago2 or Ago1 or control immunoglobin G (IgG). *P<0.05 vs IgG control. B, Smooth muscle cells (SMCs) were cocultured with bovine aortic ECs transfected with a plasmid encoding the eGPF-Ago2. The enhanced green fluorescent protein-Ago2 proteins were visualized in live ECs or SMCs by microscopy. C, The protection of miR-126 with preincubation of bovine serum albumin (BSA) or recombinant human Ago2 protein (rAgo2) for 30 minutes after RNase A digestion for 15 minutes. *P<0.05 vs miR-126 only. D, The enrichment of miR-126 in SMCs treated with miR-126 preincubated with BSA or rAgo2 for 3 hours. *P<0.05 vs miRs only. E, The levels of Ago2 in ECs (left) or EC-CM (right) under static or laminar shear stress (LSS) conditions for 24 hours. F, The levels of miR-126 in Ago2 immunocomplex in EC-CM under static of LSS conditions. Images are representative of triplicates with similar results. Semiquantification results are shown in the lower panels. *P<0.05 vs static.

Article Snippet: Human umbilical vein ECs (HUVECs) were cultured in medium 199 (Gibco) supplemented with 10% fetal bovine serum (FBS) (Omega Scientific) and 10% Endothelial Growth Medium (Cell Applications).

Techniques: Transmission Assay, Control, Transfection, Plasmid Preparation, Microscopy, Recombinant, Shear

Journal: Circulation Research

Article Title: Regulation of Vascular Smooth Muscle Cell Turnover by Endothelial Cell–Secreted MicroRNA-126

doi: 10.1161/circresaha.113.280883

Figure Lengend Snippet: Figure 6. Shear stress (SS)–imposed endothelial cells (ECs) modulate smooth muscle cell (SMC) turnover through microRNA-126 (miR-126). SMCs were monocultured or cocultured with (1) ECs kept under static condition or exposed to laminar shear stress (LSS) for 24 hours; or (2) ECs transfected with negative control inhibitor (AMC) or anti–miR-126 inhibitor (AM126). A, Proliferating cell nuclear antigen (PCNA) in the SMCs was detected by immunostaining. B, Protein levels of PCNA, cyclinA, and p21 in SMCs were assayed by Western blot. C and D, SMC proliferation and apoptosis analyzed for propidium iodide–stained DNA content and Annexin V–stained membrane flip, respectively. *P<0.05 vs Ø/SMC (SMC monocultured). #P<0.05 vs Static EC-SMC or AMC EC-SMC.

Article Snippet: Human umbilical vein ECs (HUVECs) were cultured in medium 199 (Gibco) supplemented with 10% fetal bovine serum (FBS) (Omega Scientific) and 10% Endothelial Growth Medium (Cell Applications).

Techniques: Shear, Transfection, Negative Control, Immunostaining, Western Blot, Staining, Membrane

Journal: Circulation Research

Article Title: Regulation of Vascular Smooth Muscle Cell Turnover by Endothelial Cell–Secreted MicroRNA-126

doi: 10.1161/circresaha.113.280883

Figure Lengend Snippet: Figure 7. Depletion of microRNA-126 (miR-126) in mice inhibits smooth muscle cell (SMC) proliferation and neointimal formation in arteries after cessation of blood flow. A, The miR-126 expression levels in mouse carotid arteries with or without ligation. Vascular endothelial cells (ECs) were denuded before RNA isolation. *P<0.05 vs unligated control. B, In situ hybridization of miR-126 in ligated or unligated carotid arteries from miR-126 wild-type (WT) mice. Yellow arrows, elastic lamina (EL). C, Representative images of modified Verhoeff-Van Gieson–stained ligated or unligated carotid arteries from WT or miR-126 knockout (KO) mice. Arrows indicate the thickened neointima. D and E, Representative images of immunofluorescent-stained proliferating (proliferating cell nuclear antigen [PCNA]) SMC (smooth muscle-α actin) in ligated or unligated carotid arteries from WT and KO mice. F, Representative images of modified Verhoeff-Van Gieson–stained ligated carotid arteries from WT or KO mice treated with control (CL) media or EC-conditioned media (EC-CM); or with local delivery of CL oligo or miR-126 mimics (PRE126). Arrows indicate thickened neointima. G, In situ hybridization of miR-126 in ligated carotid arteries from KO mice with treatment of EC-CM, miR-126, or the respective controls.

Article Snippet: Human umbilical vein ECs (HUVECs) were cultured in medium 199 (Gibco) supplemented with 10% fetal bovine serum (FBS) (Omega Scientific) and 10% Endothelial Growth Medium (Cell Applications).

Techniques: Expressing, Ligation, Isolation, Control, In Situ Hybridization, Modification, Staining, Knock-Out

Journal: JVS-Vascular Science

Article Title: Comparison between vascular smooth muscle cells isolated from descending thoracic and abdominal aortas of mice in cell function, phenotype features, and gene expression

doi: 10.1016/j.jvssci.2026.100411

Figure Lengend Snippet: (A) Schematic flowchart of vascular smooth muscle cell (VSMC) isolation from the thoracic and abdominal aortas of mice (figure created with BioRender.com). (B) Representative images of α-smooth muscle actin ( α-SMA ) staining (original magnification ×20) of thoracic aortic smooth muscle cell ( Th-SMC ) and abdominal aortic smooth muscle cell ( Ab-SMC ) at passages 3 and 7, respectively. Green indicates α-SMA, and blue means nuclei. (C) The ratio of α-SMA-positive cells was assessed. There was no significant difference in α-SMA expression between passages in each cell type (n = 4 per condition, two images per well, two wells analyzed). nd , not significant; P , passage.

Article Snippet: Wells were washed with phosphate-buffered saline and cultured in fetal bovine serum-free Smooth Muscle Cell Growth Medium (Basal medium, Cell Applications).

Techniques: Isolation, Staining, Expressing

Journal: JVS-Vascular Science

Article Title: Comparison between vascular smooth muscle cells isolated from descending thoracic and abdominal aortas of mice in cell function, phenotype features, and gene expression

doi: 10.1016/j.jvssci.2026.100411

Figure Lengend Snippet: Cell migration and proliferation were assessed to investigate the cell function between cell types. (A) Representative images of the scratch assay for thoracic aortic smooth muscle cells ( Th-SMCs ) and abdominal aortic smooth muscle cells ( Ab-SMCs ) at 0, 12, 24, and 72 hours. (B) Th-SMCs migrated into the scratched wound area (between the dotted lines ) significantly faster than Ab-SMCs at 24, 48, and 72 hours, as evaluated by the reduced wound region (n = 6 per group). (C) Representative immunofluorescence images of Ki67 ( red ) and 4′,6-diamidino-2-phenylindole (DAPI) ( blue ) staining during the scratch migration assay (0, 12, and 72 hours) under serum-free conditions. Scale bar , 100 μm. (D) Quantification of proliferative activity expressed as the Ki67/DAPI intensity ratio during the migration assay. Ki67/DAPI intensity was minimal and not significantly different between Th-SMCs and Ab-SMCs at any time point, indicating negligible proliferation under serum-free conditions. (E) Representative Ki67 staining images of Th-SMCs and Ab-SMCs under growth conditions at day 0, day 1, and day 4 corresponding with the proliferation assay. (F) Quantification of Ki67/DAPI intensity ratio under growth conditions, demonstrating increased proliferative activity over time and a significant difference between Th-SMCs and Ab-SMCs at day 4. ns , not significant. ∗∗∗ P ≤ .001; ∗∗∗∗ P ≤ .0001.

Article Snippet: Wells were washed with phosphate-buffered saline and cultured in fetal bovine serum-free Smooth Muscle Cell Growth Medium (Basal medium, Cell Applications).

Techniques: Migration, Cell Function Assay, Wound Healing Assay, Immunofluorescence, Staining, Activity Assay, Proliferation Assay

Journal: JVS-Vascular Science

Article Title: Comparison between vascular smooth muscle cells isolated from descending thoracic and abdominal aortas of mice in cell function, phenotype features, and gene expression

doi: 10.1016/j.jvssci.2026.100411

Figure Lengend Snippet: Oxidative stress assay on vascular smooth muscle cells (VSMCs) and expression of antioxidant markers in the oxidative stressed VSMCs. (A) VSMCs were treated with different concentrations of H 2 O 2 (0-750 μM) for 12 hours, followed by evaluating the cell viability using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Viability in thoracic aortic smooth muscle cells ( Th-SMCs ) was significantly higher than in abdominal aortic smooth muscle cells ( Ab-SMCs ) at 100 μM and 750 μM H 2 O 2 (n = 4 per group). To evaluate the expression of antioxidant markers in VSMCs treated with hydrogen peroxide ( H 2 O 2 ) at the same concentration used in the oxidative stress assay, reverse transcription-polymerase chain reaction was performed. (B) Nrf2 expression was significantly higher in Th-SMCs than in Ab-SMCs at 100 μM and 750 μM. (C, D) Similarly, Catalase and Sod1 expression were significantly greater in Th-SMCs at each concentration than in Ab-SMCs.

Article Snippet: Wells were washed with phosphate-buffered saline and cultured in fetal bovine serum-free Smooth Muscle Cell Growth Medium (Basal medium, Cell Applications).

Techniques: Expressing, Concentration Assay, Reverse Transcription, Polymerase Chain Reaction

Journal: iScience

Article Title: Targeting CDK7/12/13 functional synergism reverses myofibroblast activation and ameliorates lung fibrosis

doi: 10.1016/j.isci.2025.112778

Figure Lengend Snippet: The functional synergism of CDK7/12/13 is the critical determinant of lung myofibroblast activation (A) Heatmap showing relative expression of pro-fibrotic genes in TGFβ-induced lung myofibroblasts transfected with siRNA as indicated. Data are presented as a fold change when normalized to pro-fibrotic gene transcript levels in serum free media (SFM) starved HLFs transfected with control siRNA. (B) COL1A1 mRNA level in SFM starved HLFs and TGFβ-induced lung myofibroblasts transfected with siRNAs as indicated. (C) Representative images of αSMA immunofluorescence in SFM starved HLFs and TGFβ-induced lung myofibroblasts transfected with siRNAs as indicated. Scale bar: 50 μm. (D) Quantification of immunofluorescent intensity of αSMA in (C). (E) Representative images of a 24-well plate showing gel contraction in collagen matrices seeded with SFM starved HLFs and TGFβ-induced lung myofibroblasts transfected with siRNAs as indicated. The scale bar is the same as in (C) although not specifically indicated. (F) Quantification of collagen gel contraction in (E). Data are presented as percentage (%) of gel contraction-induced by TGFβ induced lung myofibroblasts transfected with control siRNA, which was arbitrarily set as 100%. (G) Representative western blot of nuclear RNA Pol II CTD S2P, S5P, S7P and total nuclear RNA Pol II in SFM starved HLFs and TGFβ-induced lung myofibroblasts transfected with siRNAs as indicated. Yin Yang 1 (YY1) is a loading control for nuclear extract. Data in (B), (D) and (F) represent mean ± S.E. of three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001. p value is calculated using the unpaired two-tailed t-test.

Article Snippet: For myofibroblast activation assay, after starved with serum free media (SFM) for 24h, HLFs were treated with or without 10ng/ml recombinant human TGFβ1 (R&D systems, 7754-BH) for 24 or 48h prior to terminal analyses.

Techniques: Functional Assay, Activation Assay, Expressing, Transfection, Control, Immunofluorescence, Western Blot, Two Tailed Test

Journal: iScience

Article Title: Targeting CDK7/12/13 functional synergism reverses myofibroblast activation and ameliorates lung fibrosis

doi: 10.1016/j.isci.2025.112778

Figure Lengend Snippet: CDK7/12/13 synergistically regulate lung fibroblast proliferation (A) Representative images of KI67 immunofluorescence in SFM starved or 20% FBS stimulated HLFs transfected with siRNAs as indicated. Scale bar: 50 μm. (B) Quantification of immunofluorescent intensity of KI67 in (A). (C) Proliferation of SFM starved or 20% FBS stimulated HLFs transfected with siRNAs as indicated. (D) Heatmap showing relative expression of RTKs in 20% FBS stimulated HLFs transfected with siRNAs as indicated. Data are presented as a fold change normalized to RTK transcript levels in SFM starved HLFs transfected with control siRNA. (E) MYC mRNA level in 20% FBS stimulated HLFs transfected with siRNAs as indicated. Data in (B), (C), and (E) represent mean ± S.E. of three independent experiments. ∗ p < 0.05 and ∗∗∗ p < 0.001. p value is calculated using the unpaired two-tailed t-test.

Article Snippet: For myofibroblast activation assay, after starved with serum free media (SFM) for 24h, HLFs were treated with or without 10ng/ml recombinant human TGFβ1 (R&D systems, 7754-BH) for 24 or 48h prior to terminal analyses.

Techniques: Immunofluorescence, Transfection, Expressing, Control, Two Tailed Test

Journal: iScience

Article Title: Targeting CDK7/12/13 functional synergism reverses myofibroblast activation and ameliorates lung fibrosis

doi: 10.1016/j.isci.2025.112778

Figure Lengend Snippet: Pharmacological inhibition of CDK7/12/13 by THZ1 reprograms lung myofibroblasts to fibroblasts (A) Experimental scheme of inhibition of lung myofibroblast activation by THZ1. (B) Heatmap showing TGFβ-induced differentially expressed genes (TGFβ1 plus vehicle versus SFM) in SFM starved HLFs or TGFβ-induced lung myofibroblasts treated with or without THZ1. ( p -value <0.05, FDR <0.05, log 2 -transformed fold change ≥ 1.5 or ≤ −1.5). Euclidean clustering of both rows and columns using log 2 -transformed RNA-Seq expression data, n = 3 per treatment group. (C) Volcano plot showing log 2 -transformed fold change (x axis) of TGFβ1 plus THZ1 versus TGFβ1 plus vehicle on all genes in lung myofibroblasts. Blue and red dots represent THZ1 repressed and induced genes in lung myofibroblast, respectively. (D) Gene ontology (GO) analysis (Reactome) of THZ1-induced DEGs in lung myofibroblasts. (E) Representative images of αSMA immunofluorescence in SFM starved HLFs or TGFβ-induced lung myofibroblasts treated with or without THZ1. Scale bar: 50 μm. (F) Quantification of immunofluorescent intensity of αSMA in (E). (G) Representative images of a 24-well plate showing gel contraction in collagen matrices seeded with SFM starved HLFs or TGFβ-induced lung myofibroblasts treated with or without THZ1. The scale bar is the same as in (E) although not specifically indicated. (H) Quantification of collagen gel contraction in (G). Data are presented as percentage (%) of gel contraction-induced by TGFβ-induced lung myofibroblasts treated with vehicle, which was arbitrarily set as 100%. Data in (F) and (H) represent mean ± S.E. of three independent experiments. ∗∗ p < 0.01 and ∗∗∗∗ p < 0.0001. p value is calculated using the unpaired two-tailed t-test.

Article Snippet: For myofibroblast activation assay, after starved with serum free media (SFM) for 24h, HLFs were treated with or without 10ng/ml recombinant human TGFβ1 (R&D systems, 7754-BH) for 24 or 48h prior to terminal analyses.

Techniques: Inhibition, Activation Assay, Transformation Assay, RNA Sequencing, Expressing, Immunofluorescence, Two Tailed Test

Journal: iScience

Article Title: Targeting CDK7/12/13 functional synergism reverses myofibroblast activation and ameliorates lung fibrosis

doi: 10.1016/j.isci.2025.112778

Figure Lengend Snippet: THZ1 reverses lung myofibroblast activation (A) Experimental scheme of reversion of activated lung myofibroblasts to fibroblasts by THZ1. (B) Profiling of pro-fibrotic gene expression in SFM starved HLFs and TGFβ-induced lung myofibroblasts treated with or without THZ1. (C) Representative images of αSMA immunofluorescence in SFM starved HLFs and TGFβ-induced lung myofibroblasts treated with or without THZ1. Scale bar: 50 μm. (D) Quantification of immunofluorescent intensity of αSMA in (C). (E) Representative images of a 24-well plate showing gel contraction in collagen matrices seeded with SFM starved HLFs and TGFβ-induced lung myofibroblasts treated with or without THZ1. The scale bar is the same as in (C) although not specifically indicated. (F) Quantification of collagen gel contraction in (E). Data are presented as percentage (%) of gel contraction-induced by SFM starved HLFs. Data in (B), (D), and (F) represent mean ± S.E. of three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001. p value is calculated using the unpaired two-tailed t-test.

Article Snippet: For myofibroblast activation assay, after starved with serum free media (SFM) for 24h, HLFs were treated with or without 10ng/ml recombinant human TGFβ1 (R&D systems, 7754-BH) for 24 or 48h prior to terminal analyses.

Techniques: Activation Assay, Gene Expression, Immunofluorescence, Two Tailed Test