Journal: Annals of the rheumatic diseases

Article Title: RUNX1 is expressed in a subpopulation of dermal fibroblasts and is associated with disease severity of systemic sclerosis

doi: 10.1016/j.ard.2025.10.033

Figure Lengend Snippet: TGF- β 1 increases the RUNX1 expression in SSc fibroblasts and inhibition of RUNX1 reduces ECM markers. (A) Western blot of 3 isolated fibroblasts lines treated with TGF- β 1. The blot shows all isoforms of RUNX1a, b, and c that are overexpressed under the TGF- β 1 stimulation. (B) Schematic graph illustrating the timeline for the culture and TGF- β 1 treatment of dcSSc-isolated fibroblasts, matched healthy-isolated fibroblasts, and normal human dermal (NHD) fibroblast cells. RUNX1 expression rate in samples treated with TGF- β 1 (in red) vs control (in blue) for the 24 hours after exposure. (C) Volcano plot of differentially expressed analysis of the 2 SSc-isolated fibroblast lines at 12 hours after exposure vs the baseline. (D) The pathway analysis of Reactome gene sets shows the biological pathways and processes that are significantly represented within top DEG genes of SSc-isolated fibroblast lines 12 hours after TGF- β 1 treatment vs the baseline. Data from B to D were obtained through publicly available data of GSE12493 . (E) Schematic graph showing 2 lines of SSc-isolated fibroblasts treated with siRNA against RUNX1 (siRUNX1) and nontargeting control siRNA (siNC). (F) UMAP projection and dot plot of RUNX1 and CBFB of the single-cell RNA-seq data. (G) UMAP of 10 fibroblast clusters (0–9) for siR-UNX1 and siNC. (H) Cell proportion of siRUNX1 and siNC per cluster. (I) Top 4 upregulated and downregulated marker genes per cluster. (J) Top 15 enriched pathways that are significantly represented across siRUNX1 and siNC (K) Bar plots showing the percentage of cells expressing COL1A1, FN1, COL4A1, LUM, ACTA2, LGR5, COL8A1, COMP , and THBS1 per condition (red: siRUNX1, green: siNC) or per cluster. (L) Module score for extracellular matrix organisation pathway per cluster and per condition. (M) Feature plot of the ECM module score. dcSSc, diffuse cutaneous SSc; DEG, differentially expressed gene; ECM, extracellular matrix; RUNX1, runt-related transcription factor 1; siRUNX1, siRNA targeting RUNX1; SSc, systemic sclerosis; TGF- β , transforming growth factor- β ; UMAP, uniform manifold approximation and projection.

Article Snippet: We then analysed a previously generated DNA microarray dataset (National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO): GSE12493 ) consisting of 2 independent SSc fibroblast cell lines, 1 healthy control fibroblast cell line (isolated in parallel), and 1 normal human dermal (NHD) fibroblast cell line obtained from American Type Culture Collection (ATCC), treated with 50 pM TGF- β 1 [ ] ( ).

Techniques: Expressing, Inhibition, Western Blot, Isolation, Control, Single Cell, RNA Sequencing, Marker

Journal: Annals of the rheumatic diseases

Article Title: RUNX1 is expressed in a subpopulation of dermal fibroblasts and is associated with disease severity of systemic sclerosis

doi: 10.1016/j.ard.2025.10.033

Figure Lengend Snippet: RUNX1 contributes to fibroblast activation, proliferation and contraction. (A) RUNX1 western blot of CRISPR-generated RUNX1 KO and wild-type (WT) fibroblasts under the TGF- β 1 stimulation vs control. RUNX1 isoforms of a, b, and c were marked in the blot by arrows. (B) α -SMA and RUNX1 IF staining of KO and WT fibroblasts under the TGF- β 1 stimulation vs control. (C) α -SMA western blot of KO and WT fibroblasts under the TGF- β 1 stimulation vs control. (D) ACTA2 mRNA expression of KO and WT fibroblasts under the TGF- β 1 induction vs control. (E) Fold change expression of FN1, COL1A1, LUM , and SFRP4 in TGF- β 1-induced SSc fibroblasts treated with Ro5–3335 compared to control (3 lines of SSc fibroblasts, 2 replicates each). (F) Proliferation curve of normal human dermal (NHD) fibroblasts in the presence and absence of Ro5–3335. (G,H) The 3D collagen contraction assays, fixed (G) and floating (H) models, of NHD fibroblasts treated with Ro5–3335 (4 replicates for each condition). SIS3 (SMAD3 inhibitor) was used as positive control that significantly eliminates the contraction ability of fibroblasts. Negative control is collagen matrix with no fibroblasts. The overhead pictures represent 1 replicate for each condition. (I) 3D self-assembled (SA) tissue constructs from the healthy- and SSc-isolated fibroblast lines with donors’ clinical characteristics. H&E staining of representative untreated and Ro5–3335-treated tissues. (J) Tissue area fold change of each cell line over the control for healthy and SSc SA tissues. Data from 3 healthy and 4 SSc lines, 3 replicates per line, repeated in 2 independent sets. (K) Change in area of an SSc-isolated SA tissue when treated for 1, 2, or 3 weeks with Ro5–3335 compared to control (Student’s t test P value: **.001-.01, ****<.0001 in GraphPad Prism v9). α -SMA, alpha smooth muscle actin; H&E, haematoxylin and eosin; KO, knockout; RUNX1, runt-related transcription factor 1; SSc, systemic sclerosis; TGF- β , transforming growth factor- β ;Clustered Regularly Interspaced Palindromic Repeats (CRISPR),Smad Family Member 3 (SMAD3),Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), Not Applicable (N/A), Quantitative Polymerase Chain Reaction (QPCR), Quantitative Polymerase Chain Reaction, Immunofluorescenc (IF).

Article Snippet: We then analysed a previously generated DNA microarray dataset (National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO): GSE12493 ) consisting of 2 independent SSc fibroblast cell lines, 1 healthy control fibroblast cell line (isolated in parallel), and 1 normal human dermal (NHD) fibroblast cell line obtained from American Type Culture Collection (ATCC), treated with 50 pM TGF- β 1 [ ] ( ).

Techniques: Activation Assay, Western Blot, CRISPR, Generated, Control, Staining, Expressing, Positive Control, Negative Control, Construct, Isolation, Knock-Out, Real-time Polymerase Chain Reaction

Journal: PLoS Biology

Article Title: Force-Induced Unfolding of Fibronectin in the Extracellular Matrix of Living Cells

doi: 10.1371/journal.pbio.0050268

Figure Lengend Snippet: Spatial ratiometric images and histograms of all pixels within each field of view are shown for dimeric amine/cys Fn-DA in 0 and 1 M GdnHCl and monomeric amine/cys Fn-DA in 1 and 4 M GdnHCl (A). Amine/cys Fn-DA was added to the culture medium of fibroblasts for 24 h, and excess Fn-u was added to suppress intermolecular energy transfer. Confocal microscopic images of acceptor and donor peak intensities taken 1 μm above the glass–cell interface were background subtracted, averaged, and thresholded, and the I A / I D ratiometric image of acceptor to donor was color-coded within the range of 0.05 to 1.0. A histogram (B) for all pixels of amine/cys Fn-DA–containing ECM (C) and an overlay of I A / I D on the DIC image (D) are shown in a region in which the matrix showed a transition from low to intermediate I A / I D within a single Fn fiber. Histograms are overlaid in (B) for regions of extended (E; purple) and unfolded Fn (F; pink). Histograms were generated with 0.01-ratio-unit bin widths. Scale bars = 25 μm.

Article Snippet: Primary human dermal fibroblast cells derived from foreskins (PromoCell, http://www.promocell.com/ ) were maintained for less than eight passages in Fibroblast Growth Medium plus Supplement (PromoCell).

Techniques: Generated

Journal: PLoS Biology

Article Title: Force-Induced Unfolding of Fibronectin in the Extracellular Matrix of Living Cells

doi: 10.1371/journal.pbio.0050268

Figure Lengend Snippet: Amine/cys Fn-DA and excess Fn-u were added to the culture medium of fibroblasts for 24 h. Color-coded I A / I D ratiometric images are shown for control cells (A), extracted cell-free matrix (B), and fibroblast cells after 60 min exposure to the ROCK inhibitor Y-27632 (C). Histograms with 0.01-ratio-unit bin widths for all pixels of control (black), cell-free (purple), and ROCK-inhibited matrix (pink) were derived from three random fields of view each from three separate experiments in each group (D). Solution denaturation values for dimeric Fn-DA in 0 M GdnHCl and monomeric Fn-DA in 1 and 4 M GdnHCl are shown as red, green, and blue lines, respectively. Scale bars = 50 μm.

Article Snippet: Primary human dermal fibroblast cells derived from foreskins (PromoCell, http://www.promocell.com/ ) were maintained for less than eight passages in Fibroblast Growth Medium plus Supplement (PromoCell).

Techniques: Control, Derivative Assay

Journal: PLoS Biology

Article Title: Force-Induced Unfolding of Fibronectin in the Extracellular Matrix of Living Cells

doi: 10.1371/journal.pbio.0050268

Figure Lengend Snippet: A schematic of the strain device is shown in the relaxed configuration with length L before (A) and length L + DL after (B) application of strain. PDMS sheets were covalently modified with Fn-u as described in Materials and Methods, and fibroblast cells were cultured for 24 h in the presence of amine/cys Fn-DA and excess Fn-u. Cells were extracted in mild detergent. Color-coded I A / I D ratiometric images are shown for a field of view without application of stretch (C) and after application of 70% elongation strain with 28% transverse compression (D). Region of interest analysis on individual fibrils was used to determine the impact of elongation on I A / I D on a per fibril basis (circles, mean ± standard deviation), and binned averages were calculated for fibrils between −37% and −20%, −20% and −10%, −10% and 10%, 10% and 40%, and 40% and 73% strain (red squares, mean ± standard deviations) (E). Abscissa is also plotted as relative length change. Solution values for dimeric Fn-DA in 0 M GdnHCl and monomeric Fn-DA in 1 and 4 M GdnHCl are shown as horizontal red, green, and blue lines, respectively. Scale bars = 50 μm.

Article Snippet: Primary human dermal fibroblast cells derived from foreskins (PromoCell, http://www.promocell.com/ ) were maintained for less than eight passages in Fibroblast Growth Medium plus Supplement (PromoCell).

Techniques: Modification, Cell Culture, Standard Deviation

Journal: PLoS Biology

Article Title: Force-Induced Unfolding of Fibronectin in the Extracellular Matrix of Living Cells

doi: 10.1371/journal.pbio.0050268

Figure Lengend Snippet: Cys/cys Fn-DA (A–C) or amine/cys Fn-DA (D–G) was incorporated into fibroblast matrix on Fn-u that was adsorbed to plasma cleaned PDMS, and after cell extraction the substrate was relaxed to 4/5 (A and B; 3.7% transverse stretch) or 3/5 the starting length (D–F; 10% transverse stretch). I A / I D ratiometric images of cys/cys Fn-DA–containing matrix are shown at the PDMS–ECM interface (A), where a portion of the cell-free fibers are still attached to the substrate, and from the same field of view but acquired 3 μm above the PDMS surface (B), where the strain-free Fn mat randomly diffused around its points of attachment to the underlying ECM. Histograms are shown for all pixels within the field of view at the substrate (C; black) and from the upper, strain-free confocal slice (C; pink). An I A / I D ratiometric image of amine/cys Fn-DA is shown with both detached (E) and still-attached (F) regions of matrix within the same confocal slice. Region of interest analysis was used to generate histograms (G0 for all pixels within the detached (E and G; purple) and attached (F and G; pink) regions of matrix, which were overlaid on a histogram of all pixels in the field of view (black). Scale bars = 50 μm.

Article Snippet: Primary human dermal fibroblast cells derived from foreskins (PromoCell, http://www.promocell.com/ ) were maintained for less than eight passages in Fibroblast Growth Medium plus Supplement (PromoCell).

Techniques: Clinical Proteomics, Extraction

Journal: AMB Express

Article Title: Repurposing rupatadine as topical treatment against methicillin-resistant Staphylococcus aureus

doi: 10.1186/s13568-025-01947-w

Figure Lengend Snippet: Dose–Response Curves for Cytotoxicity Testing on Human Skin Fibroblasts (HSFs). Dose–response relationships of rupatadine, DMSO (vehicle control), and doxorubicin (positive control) on HSFs cell viability. Cells were treated with increasing concentrations of test compounds for 2 h, and cell viability was assessed using the SRB colorimetric assay. IC 50 values were calculated using GraphPad Prism software. The dotted line indicates IC 50 of rupatadine (1150 µg/mL). Rupatadine demonstrated minimal cytotoxicity compared to the potent positive control doxorubicin, while DMSO showed no significant cytotoxic effects across all tested concentrations

Article Snippet: Rupatadine cytotoxicity was assessed using Sulforhodamine B (SRB) colorimetric assay on human skin fibroblasts (HSFs) (AcceGen Biotech Cat. # ABC-TC3451, USA; Obtained in September 2023).

Techniques: Control, Positive Control, Colorimetric Assay, Software

Journal: Tissue Engineering. Part C, Methods

Article Title: The Acellular Myocardial Flap: A Novel Extracellular Matrix Scaffold Enriched with Patent Microvascular Networks and Biocompatible Cell Niches

doi: 10.1089/ten.tec.2012.0536

Figure Lengend Snippet: (A) Images of decellularized porcine myocardial ECM from group 4 scaffolds seeded with rat dermal fibroblasts (FB) and cultured for either 7 (top) or 14 (bottom) days. Cells were labeled with Live/Dead reagents at each time point. Live cells were labeled with calcein AM (green) and dead cell nuclei were stained with ethidium homodimer (EthD-1, red; 20×, scale bar=50 μm). (B) Immunofluorescence (IF) images of decellularized porcine myocardial ECM from group 4 scaffolds seeded with neonatal rat cardiomyocytes (CM) and cultured for 7 days. Scaffolds were stained for α-sarcomeric actinin, cardiac MHC, actin, and connexin43, and nuclei were stained with DAPI (40×, scale bar=20 μm).

Article Snippet: For initial cytotoxicity studies, ∼5×10 6 rat dermal fibroblasts (Cell Applications) at passage 11 were seeded onto the dry scaffold samples and cultured statically for either 7 or 14 days.

Techniques: Cell Culture, Labeling, Staining, Immunofluorescence

Journal: Cell Death & Disease

Article Title: Rewiring melanoma cell fate: TRPM8 modulators trigger apoptosis and boost NK cell cytotoxicity

doi: 10.1038/s41419-026-08469-8

Figure Lengend Snippet: A Representative Western blot showing TRPM8 protein expression in the indicated cell lines. Tubulin was used as loading control. Viability of human melanocytes ( B ) and human dermal fibroblasts ( C ) untreated or treated with compounds 4 and 9 at the concentrations indicated in the legends on the right. Absorbance values from WST-1 assays at 24, 48, and 72 h are shown. Data are presented as mean ± SD of three independent experiments. n.s . indicates not significant. Representative Live/Dead assay images of human melanocytes ( D ) and human dermal fibroblasts ( E ) treated for 24 h with compounds 4 and 9 (1 or 10 μM). Viable cells are shown in green (acridine orange; total cells), while dead cells are shown in red (propidium iodide; dead cells). Overlay images are shown. Scale bar, 100 μm. Quantification of cell death is displayed to the right of each overlay image. The percentage of dead cells was calculated as: (red-stained dead cells/green-stained total cells) × 100.

Article Snippet: The Immortalized Human Dermal Fibroblasts ( P10858 -IM, Innoprot) were cultured in Fibroblast Medium-2 (FM-2; Innoprot) supplemented with 5% FBS (Gibco), 1% Fibroblast Growth Supplement-2 (FGS-2; Innoprot) and 1% penicillin-streptomycin (Gibco).

Techniques: Western Blot, Expressing, Control, Live Dead Assay, Staining

Journal: Cell reports

Article Title: TACC3 Regulates Microtubule Plus-End Dynamics and Cargo Transport in Interphase Cells

doi: 10.1016/j.celrep.2019.12.025

Figure Lengend Snippet: (A) NHDFs or SK-N-SHs treated with non-targeting (ctrl), EB1, or CLIP-170 (CLIP) siRNAs were mock-infected or infected with HSV-1 at MOI 10 for 5 h and analyzed by WB. (B) SK-N-SHs treated with the indicated siRNAs were infected, as in (A). (C) SK-N-SHs treated with independent EB2 siRNAs (I or II) were infected, as in (A). (D) SK-N-SHs were treated with 100 nM BafA or DMSO and infected at MOI 10 with VSV for 4 h or HSV-1 for 5 h. (E) NHDFs or SK-N-SHs analyzed by WB using the indicated antibodies. (F) NHDFs or SK-N-SHs stained for tyrosinated (Tyr), detyrosinated (Detyr), and acetylated (Ac) tubulin. Nuclei were stained with Hoechst. (G) NHDFs or SK-N-SHs treated with 500 nM DMSO or 10 μM nocodazole were infected at MOI 20 with HSV-1 for 4 h in the presence of 1 μg/mL actinomycin D. Fixed cells were stained for VP5 and with Hoechst. Assessed for the accumulation of VP5 over 2 biological replicates were ≥ 165 NHDF or ≥ 190 SK-N-SH nuclei; error bars, SEMs; *p < 0.05, **p < 0.01, N.S., not significant; unpaired 2-tailed t test. (H) Cells treated as in (G) were infected at MOI 10 with HSV-1 for 5 h. All of the experiments represent ≥3 replicates unless indicated. See also and .

Article Snippet: Certified Primary Normal Human Dermal Fibroblasts isolated from human male neonatal foreskin (NHDFs, Lonza CC-2509), Phoenix-AMPHO (ATCC), Vero and BSC40 cells (Dr. Ian Mohr, NYU School of Medicine) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) containing 5% Fetal Bovine Serum (FBS), 1% L-Glutamine, and 1% penicillin-streptomycin.

Techniques: Infection, Staining

Journal: Cell reports

Article Title: TACC3 Regulates Microtubule Plus-End Dynamics and Cargo Transport in Interphase Cells

doi: 10.1016/j.celrep.2019.12.025

Figure Lengend Snippet:

Article Snippet: Certified Primary Normal Human Dermal Fibroblasts isolated from human male neonatal foreskin (NHDFs, Lonza CC-2509), Phoenix-AMPHO (ATCC), Vero and BSC40 cells (Dr. Ian Mohr, NYU School of Medicine) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) containing 5% Fetal Bovine Serum (FBS), 1% L-Glutamine, and 1% penicillin-streptomycin.

Techniques: Western Blot, Virus, Retroviral, Plasmid Preparation, Recombinant, Transfection, Negative Control, Amplification, Software, Microscopy