human methylcellulose serum  (R&D Systems)

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

Journal: Cell Reports Medicine

Article Title: IL-21/IL-21R signaling renders acute myeloid leukemia stem cells more susceptible to cytarabine treatment and CAR T cell therapy

doi: 10.1016/j.xcrm.2024.101826

Figure Lengend Snippet:

Article Snippet: Human Methylcellulose Serum-Free Enriched Media , R&D Systems , Cat# HSC005SF.

Techniques: Control, Recombinant, Virus, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Plasmid Preparation, Software, Phospho-proteomics