bj cell line  (ATCC)

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