Journal: Frontiers in Oncology

Article Title: Irradiation enhances the malignancy-promoting behaviors of cancer-associated fibroblasts

doi: 10.3389/fonc.2022.965660

Figure Lengend Snippet: Effect of irradiation on the morphology, molecular markers, and proliferation of CAFs. (A, B) Morphology of CAFs without irradiation (A) and CAFs that received 8Gy irradiation (B) were compared under a light microscope (100×), and both groups of cells were spindle-shaped, but the irradiated cells were slightly disorganized; (C) Representative western blotting for FAP, α-SMA, and vimentin in CAFs that received 0Gy, 8Gy, and 15Gy irradiation; (D) The relative expression differences of the biomarkers after normalization of the GAPDH were analyzed by quantitatively comparing the density differences of the immunoblot bands; (E) MTT experiment was used to analyze the proliferation activity of CAFs under 0Gy, 8Gy, and 15Gy irradiation. T-test, * P < 0.05; (F) The sketch map of supernatants preparations of CAFs. (T-test, data are presented as the mean ± S.D. *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: The primary antibodies were: FAP (1:1000, Abclonal), Vimentin (1:50000, proteintech), and α-SMA (1:5000, proteintech).

Techniques: Irradiation, Light Microscopy, Western Blot, Expressing, Activity Assay

Journal: bioRxiv

Article Title: Monocytes Strongly Induce (MYO)Fibroblast Contraction in a New 3D Skin Model to Understand the Inflammation-Fibrosis Axis in Systemic Sclerosis

doi: 10.64898/2026.02.12.705496

Figure Lengend Snippet: ( A ) Sorted CD45-cells (fibroblasts) were analyzed for expression of myofibroblast-related genes after co-culture with monocytes in collagen hydrogels. Values (mean ± SEM) indicate relative gene expression (−ΔCt) measured with qPCR. Reference genes used were GAPDH and RPS27A. Analyses were done by two-way ANOVA with Sidak’s multiple comparisons test (**p<0.01, ***p<0.001, ****p<0.0001). ( B ) Representative immunohistochemistry staining showing robust expression of α-SMA (alpha-smooth muscle actin) (left) and fibroblast activation protein (FAP) (right) in the hydrogels containing only fibroblasts (top) compared to those co-culture with monocytes (bottom). The small blue nuclei in the bottom panels are monocytes. (Myo)fibroblasts are clearly positive for both markers (black arrows), with a typically elongated shape.

Article Snippet: Sections were then incubated with primary rabbit –anti-human alpha-smooth muscle actin (α-SMA) (1:200) (ab5694, Abcam, UK), – fibroblast activation protein (FAP) (1:100) (ab207178, Abcam), – Phospho-SMAD2 (1:100) (#3108L, CellSignaling, USA), – Phospho-Stat3 (1:50) (#9145, CellSignaling), or mouse anti-human – CD68 (#MCA1815, BioRad, USA) (1:200) antibodies, for 1 hour at room temperature.

Techniques: Expressing, Co-Culture Assay, Gene Expression, Immunohistochemistry, Staining, Activation Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting cancer-associated fibroblasts/tumor cells cross-talk inhibits intrahepatic cholangiocarcinoma progression via cell-cycle arrest

doi: 10.1186/s13046-024-03210-9

Figure Lengend Snippet: Immunofluorescence characterization of 3D-hCAFs/iCCA spheroids. ( A ). Representative confocal microscopy images of homo- and hetero-spheroids established by KKU-M213 and KKU-M156 alone or in co-culture with hCAFs. The EpCAM marker, considered an epithelial-specific marker, is shown as a green fluorescent signal. The FAP marker, used as a specific marker for fibroblast activation, is shown as a red fluorescent signal. ( B ). Positive staining for Ki-67 indicates cell proliferation and demonstrates that hCAFs influence proliferation in hetero-spheroids. Nuclei are shown as a blue fluorescent signal stained with DAPI. (20× magnification, scale bar 100 μm)

Article Snippet: Then, the spheroids were incubated with anti-FAP (1:150, Abcam, Cambridge, UK), anti-EPCAM antibodies (1:800, Cell Signaling Technologies, MA, USA) in an antibody dilution buffer (2% bovine serum albumin/0,1% Triton X-100 in TBS solution) and anti-Ki67 (1:250, Abcam, Cambridge, UK), overnight at 4 °C in rotation.

Techniques: Immunofluorescence, Confocal Microscopy, Co-Culture Assay, Marker, Activation Assay, Staining