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: International Journal of Pharmaceutics: X

Article Title: Zein-based polysaccharide-tannic acid films as multifunctional barriers to prevent post-surgical adhesions

doi: 10.1016/j.ijpx.2026.100515

Figure Lengend Snippet: In vitro characterization: A) direct cells contact onto film surfaces and CLSM images after 6 days; indirect cell proliferation after 3 and 6 days towards: B) NHDF; C) Caco-2; D) CLSM images (in blue – nuclei; in green – cytoskeleton). (mean values ± SD; n = 3), ANOVA one-way; Scheffé test (* P value <0.05; ***P value <0.001). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The in vitro characterization was carried out using normal human dermal fibroblasts (NHDF) from juvenile foreskin (PromoCell, VWR, Milan, Italy) and Human colorectal adenocarcinoma cells (Caco-2).

Techniques: In Vitro

Journal: Genes to Cells

Article Title: Derivation of human differential photoreceptor cells from adult human dermal fibroblasts by defined combinations of CRX, RAX , OTX2 and NEUROD

doi: 10.1111/gtc.12127

Figure Lengend Snippet: Induction of retina-specific genes in human dermal fibroblasts by the retroviral infection of genes for defined transcription factors. (A) RT-PCR analysis for photoreceptor-specific genes in cultured human dermal fibroblasts (NHDF) obtained from Lonza after gene transfer of several kinds of transcription factors. Recoverin, blue opsin and PDE6c genes were up-regulated by CRN transduction. ‘Negative control’: amplified water as a negative control. ‘GFP’: cultured fibroblasts after retroviral gene transfer of the GFP gene as another negative control. ‘w/o’: cultured fibroblasts without gene transfer as the other negative control. ‘SPPO’: SOX2, POU1F1, PAX6 and OTX2 . ‘SPO’: SOX2, POU1F1 and OTX2 . ‘CRN’: CRX, RAX and NEUROD . ‘Human retina’: human retinal tissue as a positive control. The amount of cDNA as a template was a half in the positive control. (B) RT-PCR analysis for photoreceptor-specific genes in cultured human dermal fibroblasts (NHDF) obtained from Promo Cell after gene transfer of several transcription factors. Recoverin, blue opsin and PDE6c genes were up-regulated by CRN or CRNO transduction. ‘w/o’: cultured fibroblasts without gene transfer as a negative control. ‘GFP’: cultured fibroblasts after retroviral gene transfer of the GFP gene as another negative control. ‘CRNO’: CRX, RAX, NEUROD and OTX2 . (C) RT-PCR analysis for photoreceptor-specific genes in cultured human dermal fibroblasts (HDF-a) obtained from ScienCell after gene transfer of several transcription factors. Recoverin, blue opsin and PDE6C genes were up-regulated by CRN or CRNO transduction. Expression levels of blue opsin were increased by additional OTX2 gene transduction. ‘w/o’: cultured fibroblasts without gene transfer as a negative control. ‘GFP’: cultured fibroblasts after retroviral gene transfer of the GFP gene as another negative control. ‘CRNO’: CRX , RAX , NEUROD and OTX2 . ‘1’, ‘2’ and ‘3’ mean independently cultured, transfected and harvested cells by the same combination of CRN genes. (D) Immunocytochemistry using antibodies to rhodopsin and blue opsin (green). Nuclei were stained with DAPI (blue). Experiments were carried out at 2 weeks after infection. The cells in the left panel and the right panel are CRN-infected Fib#2 and Fib#1, respectively. Scale bars represent 10 μm.

Article Snippet: Three strains of cultured human dermal fibroblasts were used: one was obtained from Lonza (NHDF), another was from Promo Cell (NHDF) and the other was from ScienCell (HDF-a).

Techniques: Infection, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Transduction, Negative Control, Amplification, Positive Control, Expressing, Transfection, Immunocytochemistry, Staining

Journal: Oncotarget

Article Title: Evaluation of an Actinomycin D/VX-680 aurora kinase inhibitor combination in p53-based cyclotherapy

doi:

Figure Lengend Snippet: (A) Pretreatment with low dose ActD decreases the proportion of 8N cells caused by VX-680 in NHDF cells. ActD was added 24 hours post-seeding. VX-680 was added 48 hours post-seeding. Cells were harvested 96 hours after seeding. (B) NHDFs were treated with 4 nM ActD for 72 hours, fixed and stained with Giemsa (top panel). In the bottom panel, ActD was removed from the medium and cells were allowed to proliferate for 6 days before Giemsa staining.

Article Snippet: NHDF cells were bought from Promocell.

Techniques: Staining

Journal: Oncotarget

Article Title: Evaluation of an Actinomycin D/VX-680 aurora kinase inhibitor combination in p53-based cyclotherapy

doi:

Figure Lengend Snippet: (A and B) NHDF cells were treated for 48 hours with VX-680 alone (labelled as no preinc.), or treated for 48 hours with VX-680 after preincubating with the indicated amounts of ActD for 24 hours. Thereafter cells were fixed, stained Giemsa (A) and counted (B). Arrows in (A) indicate cells with abnormal nuclei. (C and D) NHDF cells treated as in A and B were allowed to recover for 6 days in drug-free medium. The growth of the cultures in drug-free medium is quantified in panel (C) by counting the total number of cells per field before and after the recovery period. The average number of cells per field with aberrant nuclei before and after the recovery period is shown in panel (D). Error bars correspond to standard deviations.

Article Snippet: NHDF cells were bought from Promocell.

Techniques: Staining

Journal: Oncotarget

Article Title: Evaluation of an Actinomycin D/VX-680 aurora kinase inhibitor combination in p53-based cyclotherapy

doi:

Figure Lengend Snippet: (A) NHDF cells were treated with the indicated amounts of ActD for 4 or 12 hours. p53 and p21 were analysed by Western blotting. α-tubulin was detected as a loading control. (B) Example of nuclear abnormalities in NHDF cells expressing a dominant negative form of p53. The percentage of cells with this sort of aberrant nuclei was 2.5 fold higher than in the corresponding cells with intact p53. Severely damaged cells, such as the one in the center of this picture, only appeared in the fibroblasts with dominant negative p53.

Article Snippet: NHDF cells were bought from Promocell.

Techniques: Western Blot, Expressing, Dominant Negative Mutation

Journal:

Article Title: ?-Melanocyte-Stimulating Hormone Counteracts the Suppressive Effect of UVB on Nrf2 and Nrf-Dependent Gene Expression in Human Skin

doi: 10.1210/en.2008-1315

Figure Lengend Snippet: Expression analysis of Nrf1-3 in human skin cells in vitro and in situ. A, Detection of Nrf1-3 in various human skin cell types in vitro as shown by conventional RT-PCR analysis. NC, Negative control, H2O as template; 1, NHK; 2, HaCaT; 3, NHM; 4, HDF; 5, HMEC-1. B, Protein expression of Nrf2 in human skin cell types. Total cell lysates (50 μg/lane) were subjected to Western immunoblot analysis using an anti-Nrf2 antibody. The same blot was reprobed with an anti-α-tubulin antibody. C, Expression of Nrf2 in healthy human skin. Nrf2 was detected by immunohistochemistry using the immunoperoxidase technique (a). Note specific cytoplasmic Nrf2 immunoreactivity within the basal layer of epidermal keratinocytes (red cells, arrows) but not in the IgG control (c). Double immunostaining of Nrf2 and melanocytes by the immunoperoxidase (Nrf2) and immunogold-silver enhancement technique (melanocytes) using a pan-Mel antibody (b). Note gray-blackish precipitate within a melanocyte but lack of Nrf2 staining within (arrow).

Article Snippet: Cell culture Normal human keratinocytes (NHK) and human dermal fibroblasts (HDF) were all derived from newborn foreskin and purchased from PromoCell (Heidelberg, Germany).

Techniques: Expressing, In Vitro, In Situ, Reverse Transcription Polymerase Chain Reaction, Negative Control, Western Blot, Immunohistochemistry, Double Immunostaining, 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