human foreskin fibroblasts hff  (ATCC)

Journal: Neoplasia (New York, N.Y.)

Article Title: Cancer-associated fibroblast-derived protein S100-A11 influences the response to anti-HER2 therapies in HER2-positive breast cancer

doi: 10.1016/j.neo.2026.101318

Figure Lengend Snippet: Characterization of CAF-200 fibroblasts and their functional impact on anti-HER2 therapy response. A. Protein expression analysis of fibroblast-associated markers in CAF-200 and normal fibroblasts (NF-31 and NF-39) by WB. CAF-200 displayed a CAF-like phenotype, with differential expression of Snail, αSMA, and caveolin-1 compared with normal fibroblasts. β-actin was used as a loading control. B. Functional impact of CM on BCCL response to anti-HER2 therapy. CM was collected from CAF-200 or NF-39 fibroblasts cultured under basal (untreated) conditions and subsequently applied to tumour cells in combination with TP. CM derived from CAF-200 reduced sensitivity to TP treatment, whereas CM from NF-39 did not reproduce this resistance-promoting effect. Cell proliferation was assessed after 5 days of treatment. Data are presented as mean ± SD (n ≥ 3). Statistical significance is indicated as follows: (*) p < 0.05; (**) p < 0.01; (***) p < 0.001. C. Effect of exogenous addition of S100-A11r, ranging from 1 to 40 ng/ml, on proliferation rates of BT-474, EFM-192A, AU-565, and SK-BR-3 cells. D. WB analysis of phosphorylated and total forms of STAT3, AKT, and ERK proteins for BT-474 and EFM-192A cells, respectively. The effect of exogenous addition of recombinant S100-A11 was assessed for 6 h.

Article Snippet: The presence of S100-A11 in CM from CAF-200 fibroblasts was quantified using the RayBio Human S100-A11 ELISA kit (RayBiotech, Peachtree Corners, GA, USA).

Techniques: Functional Assay, Expressing, Quantitative Proteomics, Control, Cell Culture, Derivative Assay, Recombinant

Journal: Neoplasia (New York, N.Y.)

Article Title: Cancer-associated fibroblast-derived protein S100-A11 influences the response to anti-HER2 therapies in HER2-positive breast cancer

doi: 10.1016/j.neo.2026.101318

Figure Lengend Snippet: The impact of TPD therapy on HER2+ breast cancer cells is contingent on the presence of S100-A11 in the CM secreted by CAF-200. A. The initial reduction in proliferation rates in the BT-474 cell line by TPD was attenuated by the addition of CAF-200–CM; however, this effect was less pronounced after S100A11 gene silencing in CAF-200 fibroblasts. CM [CAF-200] was obtained from CAF-200 treated with TPD under the indicated conditions (siC or siS100A11). The same effect was observed in the EFM-192A cell line. Treatment for 5 days with TPD therapy (15 μg/ml T; 20 μg/ml P; 0.5 and 1 nM D, respectively). Basal: control without recombinant protein; CM [CAF-200]: CAF-200–CM; siC: control silencing; si S100A11 : silencing of S100A11 gene. (*): p < 0.05; (**): p < 0.01; (***): p < 0.001. Error bars represent the calculated value of the standard deviation (n = 6). B. WB analysis of BT-474 and EFM-192A cells, respectively, of phosphorylated and total forms of STAT3, AKT, and ERK proteins. The effect of the CAF-200–CM obtained after S100A11 gene silencing in CAF-200 fibroblasts was assessed for 6 h. Relative abundance levels of up- or down-regulated proteins were determined by densitometric analysis of the images, normalising them to β-actin loading control and to the respective untreated control. All immunoblot comparisons were performed within the same membrane and exposure conditions; therefore, signal intensities should not be compared across different figures. Representative images are shown for n = 3. Other specific details of the experimental conditions are shown in the legend, unless otherwise indicated.

Article Snippet: The presence of S100-A11 in CM from CAF-200 fibroblasts was quantified using the RayBio Human S100-A11 ELISA kit (RayBiotech, Peachtree Corners, GA, USA).

Techniques: Control, Recombinant, Standard Deviation, Western Blot, Membrane