dermal fibroblast hdf cell lines  (ATCC)

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: Communications Biology

Article Title: Intracellular complement Factor H promotes tumor progression through modulation of cell cycle and actin cytoskeleton

doi: 10.1038/s42003-026-09807-4

Figure Lengend Snippet: A Progression-Free Survival Kaplan–Meier curves according to CFH expression in the KIRC TCGA cohort ( n = 508). B UMAP displaying CFH expression across labelled Seurat clusters from 20 different ccRCC primary tumors . C Dot plot displaying CFH scaled mean expression levels (dot color) and percentage of cells (dot size) within each Seurat cluster. D Hematoxylin-eosin (H&E) section of a representative primary ccRCC tumor used for spatial transcriptomic analysis (left); Same section displaying labelled spots for CFH and fibroblast signature high and low categories (middle), or CA9 high and low categories (right). Scale bar 1 mm. E Immunofluorescence of CA9 (pink), alpha-SMA (orange) and Factor H (white) of a ccRCC primary tumor. F alpha-SMA (orange) positive CAF presenting positive FH staining (white). G ccRCC cancer cells positive for CA9 (pink) and for FH (white) staining. F , G Nuclei are stained with DAPI (blue).

Article Snippet: Primary Normal Human Dermal Fibroblasts (NHDF, Promocell C-12302), the human dermal fibroblast cell line BJ (ATCC CRL-2522) and two ccRCC cell lines (A498 and Caki-1, ATCC HTB-44 & HTB-46, respectively, both p53 wild type, the first being VHL mutated, but not the second one) were employed.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Communications Biology

Article Title: Intracellular complement Factor H promotes tumor progression through modulation of cell cycle and actin cytoskeleton

doi: 10.1038/s42003-026-09807-4

Figure Lengend Snippet: A FH western blot with plasma purified FH along with lysates and SN from A498 and Caki-1 ccRCC cells (left and center, respectively), and BJ fibroblasts (right). B Western blot FH gene silencing validation in lysates from A498 cell (left), Caki-1 cells (center left), BJ fibroblasts (center right) and NHDF primary fibroblasts (right). C Western blot FH gene silencing validation in SN from A498 cell (left), Caki-1 cells (center left), BJ fibroblasts (center right) and NHDF primary fibroblasts (right). D FH quantification by ELISA on the different subcellular BJ fibroblasts’ fractions. Average +/- SD, all data points are presented. E FH and compartment-specific proteins western blot for the different cell fractions in BJ fibroblasts (left), NHDF fibroblasts (center left), A498 cells (center right) and Caki-1 cells (right). Cyto = cytosol fraction, Orga = organelle fraction and Nuc = nuclear fraction. All gel images represent bands from the same experimental run, with separation into two boxes when unrelated intermediate lanes were present in the original blot. F – H Partial co-localization of FH with the nuclear staining: F Confocal microscopy evidencing staining for FH (red, rabbit anti-FH polyclonal, ProteinTech), the nuclear staining (DAPI, blue) and the actin cytoskeleton (phalloidin, green) in A498. G Chromogen staining by immunohistochemistry for FH (Ox24 monoclonal anti-FH, brown) and nuclei (blue) of a section of a ccRCC patient tumor. Scale bar of the main image – 100 µm and of the insert, 50 µm. H Immunofluorescent staining for FH (red, rabbit anti-FH polyclonal, ProteinTech), tumor cells (CA9, white) and nuclei (DAPI) in a section of a ccRCC patient tumor. G , H The inserts represent a zoomed image.

Article Snippet: Primary Normal Human Dermal Fibroblasts (NHDF, Promocell C-12302), the human dermal fibroblast cell line BJ (ATCC CRL-2522) and two ccRCC cell lines (A498 and Caki-1, ATCC HTB-44 & HTB-46, respectively, both p53 wild type, the first being VHL mutated, but not the second one) were employed.

Techniques: Western Blot, Clinical Proteomics, Purification, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Staining, Confocal Microscopy, Immunohistochemistry

Journal: Communications Biology

Article Title: Intracellular complement Factor H promotes tumor progression through modulation of cell cycle and actin cytoskeleton

doi: 10.1038/s42003-026-09807-4

Figure Lengend Snippet: A Protein-protein interaction network analysis of common FH interacting proteins within A498 ccRCC cells and BJ fibroblasts (FH immunoprecipitated with anti-C-terminal FH mAb C18/3); STRING Tool, physical subnetwork with at least 0.4 confidence. Top 10 gene-ontology pathways for B the common FH interactors between A498 and BJ cells, C FH interacting candidates in BJ fibroblasts and D FH interactors identified in A498 ccRCC cells. E Protein-protein interaction ELISA dose-response analysis of purified FH binding to intracellular FH interacting candidates. F Validation of the co-immunoprecipitation of E2F3 and CAPZB together with FH. A498 lysate was incubated with anti-C-terminus FH mAb C18/3 (FH) or unspecific isotype control immunoglobuling G (IgG). Immunoprecipitation was then performed on protein A/G beads and proteins were eluted and probed by western blot for FH (upper line), E2F3 (middle line) and CAPZB (lower line). The validation of the target is based on its presence in the immunoprecipitates fraction with anti-FH (ip FH) but not in the one of the control IgG (ip IgG). Input: starting lysate material; SN: supernatant; ip: immunoprecipitates. G Kinetic SPR analysis using purified FH at concentrations ranging from 31.25 nM to 500 nM, twofold dilutions. FH was injected in normal (left) or low (right) ionic strength conditions on its respective FH interacting candidate coated chip for 240 s followed by 240 s dissociation. A 1:1 interaction with a drifting baseline curve was fitted to calculate kinetic parameters. The straight line represents the measured signal. The dotted one represents the kinetic fit. Curves from high to low RU values match the FH concentrations used (high FH, high RU values).

Article Snippet: Primary Normal Human Dermal Fibroblasts (NHDF, Promocell C-12302), the human dermal fibroblast cell line BJ (ATCC CRL-2522) and two ccRCC cell lines (A498 and Caki-1, ATCC HTB-44 & HTB-46, respectively, both p53 wild type, the first being VHL mutated, but not the second one) were employed.

Techniques: Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Purification, Binding Assay, Biomarker Discovery, Incubation, Control, Western Blot, Injection

Journal: Communications Biology

Article Title: Intracellular complement Factor H promotes tumor progression through modulation of cell cycle and actin cytoskeleton

doi: 10.1038/s42003-026-09807-4

Figure Lengend Snippet: A IPA comparison analysis of common predicted upstream regulators in siFH vs siC treated A498, Caki-1 and BJ cells. Top 3 upstream regulators across cell lines are annotated and a heatmap with their activation z-score in siFH is presented. B Protein-protein interaction network analysis of common FH interacting proteins and p53; STRING Tool, physical subnetwork with at least 0.4 confidence. C TP53 expression fold change (siFH/siC) for A498, Caki-1 and BJ cells. GSEA plots for the p53 pathway (hallmark gene set) comparing siFH vs siC treated D A498 ccRCC cells, E Caki-1 ccRCC cells and F BJ fibroblasts. Western blot analysis of G total p53 and H phosphorylated p53 S46 levels in the lysates of siC and siFH treated A498 ccRCC cells, Caki-1 ccRCC cells and BJ fibroblasts. I Immunofluorescence staining of p53 (light yellow) and nuclei (blue) of siC (left), siTP53 (left center), siFH (right center) or siTP53 + siFH (right) treated A498 cells (top row) and BJ fibroblasts (bottom row). Arrows indicate representative nuclei positive for p53 staining. The scale bar in the insert is of 100 µm and of the main image – 500 µm. J Evaluation of proliferation (left panels) and mortality (right panels) of siC, siTP53, siFH and siTP53 + siFH treated A498 (top row) and BJ (bottom row) cells. Cell proliferation is shown as inversed Fold Change of CFSE geometric means and mortality is represented as the Fold Change of DAPI stained dead cells. Exact p values indicated on the figures. Brown–Forsythe and Welch ANOVA tests plus post-hoc Tamhane’s T2 multiple comparisons test.

Article Snippet: Primary Normal Human Dermal Fibroblasts (NHDF, Promocell C-12302), the human dermal fibroblast cell line BJ (ATCC CRL-2522) and two ccRCC cell lines (A498 and Caki-1, ATCC HTB-44 & HTB-46, respectively, both p53 wild type, the first being VHL mutated, but not the second one) were employed.

Techniques: Comparison, Activation Assay, Expressing, Western Blot, Immunofluorescence, Staining

Journal: Communications Biology

Article Title: Intracellular complement Factor H promotes tumor progression through modulation of cell cycle and actin cytoskeleton

doi: 10.1038/s42003-026-09807-4

Figure Lengend Snippet: UMAP displaying A the malignant cell states identified by reclustering, and B the proximal tubular cell signature expression across malignant cells. C Dot plot displaying significantly enriched pathways across malignant cell states, scaled mean expression levels (dot color) and percentage of cells (dot size). D Dot plot displaying CFH scaled mean expression levels (dot color) across malignant cell states. E Pseudotime trajectories of malignant cells inferred by Monocle3. Two main branches were identified: Branch 1 representing cells undergoing cell cycle progression, and Branch 2 representing cancer cell differentiation. F Expression of E2F target genes along the trajectory of Branch 1. G Expression of CFH , POLA1 , CCND2 and MKI67 genes along the trajectory of Branch 1. H Immunofluorescence of CA9 (pink), Factor H (white) and Ki67 (light blue) of a ccRCC primary tumor. Representative Ki67 and FH positive (top row), and Ki67 and FH negative (bottom row) cancer cell staining. I Evaluation of Ki67 positive cell frequency in FH positive versus negative ccRCC cancer cells. The exact p value is indicated on the figure. Wilcoxon matched-pairs signed rank test. J Overall Survival Kaplan–Meier curves according to Hedgehog cancer cell signature abundance in the KIRC TCGA cohort ( n = 508). K Proposed mode of action of intracellular FH in ccRCC fibroblasts and cancer cells. Created in BioRender. Roumenina, L. (2025) https://BioRender.com/gtfgxgw .

Article Snippet: Primary Normal Human Dermal Fibroblasts (NHDF, Promocell C-12302), the human dermal fibroblast cell line BJ (ATCC CRL-2522) and two ccRCC cell lines (A498 and Caki-1, ATCC HTB-44 & HTB-46, respectively, both p53 wild type, the first being VHL mutated, but not the second one) were employed.

Techniques: Expressing, Cell Differentiation, Immunofluorescence, Staining

Journal: Communications Biology

Article Title: Intracellular complement Factor H promotes tumor progression through modulation of cell cycle and actin cytoskeleton

doi: 10.1038/s42003-026-09807-4

Figure Lengend Snippet: A Progression-Free Survival Kaplan–Meier curves according to CFH expression in the KIRC TCGA cohort ( n = 508). B UMAP displaying CFH expression across labelled Seurat clusters from 20 different ccRCC primary tumors . C Dot plot displaying CFH scaled mean expression levels (dot color) and percentage of cells (dot size) within each Seurat cluster. D Hematoxylin-eosin (H&E) section of a representative primary ccRCC tumor used for spatial transcriptomic analysis (left); Same section displaying labelled spots for CFH and fibroblast signature high and low categories (middle), or CA9 high and low categories (right). Scale bar 1 mm. E Immunofluorescence of CA9 (pink), alpha-SMA (orange) and Factor H (white) of a ccRCC primary tumor. F alpha-SMA (orange) positive CAF presenting positive FH staining (white). G ccRCC cancer cells positive for CA9 (pink) and for FH (white) staining. F , G Nuclei are stained with DAPI (blue).

Article Snippet: Primary Normal Human Dermal Fibroblasts (NHDF, Promocell C-12302), the human dermal fibroblast cell line BJ (ATCC CRL-2522) and two ccRCC cell lines (A498 and Caki-1, ATCC HTB-44 & HTB-46, respectively, both p53 wild type, the first being VHL mutated, but not the second one) were employed.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Communications Biology

Article Title: Intracellular complement Factor H promotes tumor progression through modulation of cell cycle and actin cytoskeleton

doi: 10.1038/s42003-026-09807-4

Figure Lengend Snippet: A FH western blot with plasma purified FH along with lysates and SN from A498 and Caki-1 ccRCC cells (left and center, respectively), and BJ fibroblasts (right). B Western blot FH gene silencing validation in lysates from A498 cell (left), Caki-1 cells (center left), BJ fibroblasts (center right) and NHDF primary fibroblasts (right). C Western blot FH gene silencing validation in SN from A498 cell (left), Caki-1 cells (center left), BJ fibroblasts (center right) and NHDF primary fibroblasts (right). D FH quantification by ELISA on the different subcellular BJ fibroblasts’ fractions. Average +/- SD, all data points are presented. E FH and compartment-specific proteins western blot for the different cell fractions in BJ fibroblasts (left), NHDF fibroblasts (center left), A498 cells (center right) and Caki-1 cells (right). Cyto = cytosol fraction, Orga = organelle fraction and Nuc = nuclear fraction. All gel images represent bands from the same experimental run, with separation into two boxes when unrelated intermediate lanes were present in the original blot. F – H Partial co-localization of FH with the nuclear staining: F Confocal microscopy evidencing staining for FH (red, rabbit anti-FH polyclonal, ProteinTech), the nuclear staining (DAPI, blue) and the actin cytoskeleton (phalloidin, green) in A498. G Chromogen staining by immunohistochemistry for FH (Ox24 monoclonal anti-FH, brown) and nuclei (blue) of a section of a ccRCC patient tumor. Scale bar of the main image – 100 µm and of the insert, 50 µm. H Immunofluorescent staining for FH (red, rabbit anti-FH polyclonal, ProteinTech), tumor cells (CA9, white) and nuclei (DAPI) in a section of a ccRCC patient tumor. G , H The inserts represent a zoomed image.

Article Snippet: Primary Normal Human Dermal Fibroblasts (NHDF, Promocell C-12302), the human dermal fibroblast cell line BJ (ATCC CRL-2522) and two ccRCC cell lines (A498 and Caki-1, ATCC HTB-44 & HTB-46, respectively, both p53 wild type, the first being VHL mutated, but not the second one) were employed.

Techniques: Western Blot, Clinical Proteomics, Purification, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Staining, Confocal Microscopy, Immunohistochemistry

Journal: Communications Biology

Article Title: Intracellular complement Factor H promotes tumor progression through modulation of cell cycle and actin cytoskeleton

doi: 10.1038/s42003-026-09807-4

Figure Lengend Snippet: A Protein-protein interaction network analysis of common FH interacting proteins within A498 ccRCC cells and BJ fibroblasts (FH immunoprecipitated with anti-C-terminal FH mAb C18/3); STRING Tool, physical subnetwork with at least 0.4 confidence. Top 10 gene-ontology pathways for B the common FH interactors between A498 and BJ cells, C FH interacting candidates in BJ fibroblasts and D FH interactors identified in A498 ccRCC cells. E Protein-protein interaction ELISA dose-response analysis of purified FH binding to intracellular FH interacting candidates. F Validation of the co-immunoprecipitation of E2F3 and CAPZB together with FH. A498 lysate was incubated with anti-C-terminus FH mAb C18/3 (FH) or unspecific isotype control immunoglobuling G (IgG). Immunoprecipitation was then performed on protein A/G beads and proteins were eluted and probed by western blot for FH (upper line), E2F3 (middle line) and CAPZB (lower line). The validation of the target is based on its presence in the immunoprecipitates fraction with anti-FH (ip FH) but not in the one of the control IgG (ip IgG). Input: starting lysate material; SN: supernatant; ip: immunoprecipitates. G Kinetic SPR analysis using purified FH at concentrations ranging from 31.25 nM to 500 nM, twofold dilutions. FH was injected in normal (left) or low (right) ionic strength conditions on its respective FH interacting candidate coated chip for 240 s followed by 240 s dissociation. A 1:1 interaction with a drifting baseline curve was fitted to calculate kinetic parameters. The straight line represents the measured signal. The dotted one represents the kinetic fit. Curves from high to low RU values match the FH concentrations used (high FH, high RU values).

Article Snippet: Primary Normal Human Dermal Fibroblasts (NHDF, Promocell C-12302), the human dermal fibroblast cell line BJ (ATCC CRL-2522) and two ccRCC cell lines (A498 and Caki-1, ATCC HTB-44 & HTB-46, respectively, both p53 wild type, the first being VHL mutated, but not the second one) were employed.

Techniques: Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Purification, Binding Assay, Biomarker Discovery, Incubation, Control, Western Blot, Injection

Journal: Communications Biology

Article Title: Intracellular complement Factor H promotes tumor progression through modulation of cell cycle and actin cytoskeleton

doi: 10.1038/s42003-026-09807-4

Figure Lengend Snippet: A IPA comparison analysis of common predicted upstream regulators in siFH vs siC treated A498, Caki-1 and BJ cells. Top 3 upstream regulators across cell lines are annotated and a heatmap with their activation z-score in siFH is presented. B Protein-protein interaction network analysis of common FH interacting proteins and p53; STRING Tool, physical subnetwork with at least 0.4 confidence. C TP53 expression fold change (siFH/siC) for A498, Caki-1 and BJ cells. GSEA plots for the p53 pathway (hallmark gene set) comparing siFH vs siC treated D A498 ccRCC cells, E Caki-1 ccRCC cells and F BJ fibroblasts. Western blot analysis of G total p53 and H phosphorylated p53 S46 levels in the lysates of siC and siFH treated A498 ccRCC cells, Caki-1 ccRCC cells and BJ fibroblasts. I Immunofluorescence staining of p53 (light yellow) and nuclei (blue) of siC (left), siTP53 (left center), siFH (right center) or siTP53 + siFH (right) treated A498 cells (top row) and BJ fibroblasts (bottom row). Arrows indicate representative nuclei positive for p53 staining. The scale bar in the insert is of 100 µm and of the main image – 500 µm. J Evaluation of proliferation (left panels) and mortality (right panels) of siC, siTP53, siFH and siTP53 + siFH treated A498 (top row) and BJ (bottom row) cells. Cell proliferation is shown as inversed Fold Change of CFSE geometric means and mortality is represented as the Fold Change of DAPI stained dead cells. Exact p values indicated on the figures. Brown–Forsythe and Welch ANOVA tests plus post-hoc Tamhane’s T2 multiple comparisons test.

Article Snippet: Primary Normal Human Dermal Fibroblasts (NHDF, Promocell C-12302), the human dermal fibroblast cell line BJ (ATCC CRL-2522) and two ccRCC cell lines (A498 and Caki-1, ATCC HTB-44 & HTB-46, respectively, both p53 wild type, the first being VHL mutated, but not the second one) were employed.

Techniques: Comparison, Activation Assay, Expressing, Western Blot, Immunofluorescence, Staining

Journal: Communications Biology

Article Title: Intracellular complement Factor H promotes tumor progression through modulation of cell cycle and actin cytoskeleton

doi: 10.1038/s42003-026-09807-4

Figure Lengend Snippet: UMAP displaying A the malignant cell states identified by reclustering, and B the proximal tubular cell signature expression across malignant cells. C Dot plot displaying significantly enriched pathways across malignant cell states, scaled mean expression levels (dot color) and percentage of cells (dot size). D Dot plot displaying CFH scaled mean expression levels (dot color) across malignant cell states. E Pseudotime trajectories of malignant cells inferred by Monocle3. Two main branches were identified: Branch 1 representing cells undergoing cell cycle progression, and Branch 2 representing cancer cell differentiation. F Expression of E2F target genes along the trajectory of Branch 1. G Expression of CFH , POLA1 , CCND2 and MKI67 genes along the trajectory of Branch 1. H Immunofluorescence of CA9 (pink), Factor H (white) and Ki67 (light blue) of a ccRCC primary tumor. Representative Ki67 and FH positive (top row), and Ki67 and FH negative (bottom row) cancer cell staining. I Evaluation of Ki67 positive cell frequency in FH positive versus negative ccRCC cancer cells. The exact p value is indicated on the figure. Wilcoxon matched-pairs signed rank test. J Overall Survival Kaplan–Meier curves according to Hedgehog cancer cell signature abundance in the KIRC TCGA cohort ( n = 508). K Proposed mode of action of intracellular FH in ccRCC fibroblasts and cancer cells. Created in BioRender. Roumenina, L. (2025) https://BioRender.com/gtfgxgw .

Article Snippet: Primary Normal Human Dermal Fibroblasts (NHDF, Promocell C-12302), the human dermal fibroblast cell line BJ (ATCC CRL-2522) and two ccRCC cell lines (A498 and Caki-1, ATCC HTB-44 & HTB-46, respectively, both p53 wild type, the first being VHL mutated, but not the second one) were employed.

Techniques: Expressing, Cell Differentiation, Immunofluorescence, Staining

Journal: International Microbiology

Article Title: Myconanotechnology: evaluation of Ag-Cu bimetallic nanoparticles synthesized by Ganoderma aff. australe against pathogens and cancer cells

doi: 10.1007/s10123-026-00793-5

Figure Lengend Snippet: Cytotoxic activity of Ganoderma aff. australe aqueous extract and Ag/Cu nanoparticles against cancer and non-cancerous cell lines evaluated by MTT assay. Cells (4,500 cells/well) were seeded in 96-well plates and allowed to adhere for 24 h before treatment. Concentrations tested were based on the amount of aqueous extract used to synthesize nanoparticles (see Table 1 ). All treatments were incubated with cells for 72 h at 37 °C with 5% CO₂. After incubation, cells were rinsed with PBS and incubated with 10 µl MTT solution (5 mg/ml) for 4 h, followed by addition of 100 µl DMSO. Absorbance was measured at 570 nm. Bar graphs show cell viability (expressed as IC₅₀ in mg/ml equivalent of extract) for five cancer cell lines and two non-cancerous control lines. Cancer cell lines: Caco-2 (colon cancer, ATCC HTB-37), HT-29 (colon cancer, ATCC HTN-38), MCF7 (breast cancer, ATCC HTB-22), A-172 (glioblastoma, ATCC CRL-1620), and U-87 MG (glioblastoma, ATCC HTB-14). Non-cancerous control lines: HDFn (human dermal fibroblasts, ATCC PCS-201-010) and Detroit 551 (normal skin fibroblasts, ATCC CCL-110). All cell lines were cultured in DMEM/F12 medium supplemented with 10% FBS, 1% antibiotic-antimycotic, 1% glutamine, 1% nonessential amino acids, and 1% sodium pyruvate. A Aqueous extract (0.5 g/50 ml) showing moderate cytotoxic activity with IC₅₀ values ranging from 1.61 ± 0.35 mg/ml (Caco-2) to 5.78 ± 1.48 mg/ml (Detroit 551), demonstrating baseline bioactivity of fungal metabolites. B M2-3-3 nanoparticles (2 ml extract + 3 ml AgNO₃ + 3 ml CuSO₄) exhibiting the highest cytotoxic efficacy across all cancer cell lines, with particularly remarkable activity against glioblastoma lines A-172 (IC₅₀: 0.26 ± 0.09 mg/ml) and U-87 MG (IC₅₀: 0.31 ± 0.12 mg/ml), and colorectal cancer lines Caco-2 (IC₅₀: 0.39 ± 0.12 mg/ml) and HT-29 (IC₅₀: 0.58 ± 0.28 mg/ml). Critically, M2-3-3 showed selective cytotoxicity with significantly higher IC₅₀ values in non-cancerous lines HDFn (2.87 ± 0.64 mg/ml) and Detroit 551 (3.45 ± 0.89 mg/ml), indicating preferential toxicity toward cancer cells. C M3-2-2 nanoparticles (3 ml extract + 2 ml AgNO₃ + 2 ml CuSO₄) demonstrating intermediate cytotoxic activity with IC₅₀ values consistently higher than M2-3-3 but lower than M5-3-3 across all cancer cell lines. D M5-3-3 nanoparticles (5 ml extract + 3 ml AgNO₃ + 3 ml CuSO₄) showing the lowest cytotoxic activity among the three nanoformulations, though still superior to the crude extract. Data represent mean ± SD of three independent experiments performed in triplicate. Statistical analysis performed using one-way ANOVA followed by Tukey’s HSD post-hoc test. Asterisks indicate significant differences: * p ≤ 0.050, ** p ≤ 0.010, *** p ≤ 0.001. The hierarchical efficacy pattern (M2-3-3 > M3-2-2 > M5-3-3 > crude extract) demonstrates successful nanotechnological enhancement of anticancer properties and establishes M2-3-3 as the optimal formulation with superior therapeutic selectivity. The exceptional sensitivity of glioblastoma cell lines suggests potential application in treating brain cancers, which are notoriously difficult to treat due to blood-brain barrier penetration challenges. Complete IC₅₀ values and statistical comparisons are provided in Table 4

Article Snippet: Fibroblast cell lines HDFn and Detroit 551 (ATCC, PCS-201-010 and CCL-110, respectively) were used to control non-cancerous cells.

Techniques: Activity Assay, MTT Assay, Incubation, Control, Cell Culture, Formulation