canine ec basal medium  (Cell Applications Inc)

Journal: Cell Death & Disease

Article Title: MCL-1 as a molecular switch between myofibroblastic and pro-angiogenic features of breast cancer-associated fibroblasts

doi: 10.1038/s41419-025-07920-6

Figure Lengend Snippet: A qRT-PCR of VEGF-A, FGF2, and ANGPT1 mRNA in bCAFsgCTRL or sgMCL-1 normalized on RPLPO mRNA expression. Mean and SEM of five independent experiments are represented as relative quantity of mRNA. Student t -test, *** P < 0.001, ns not significant. B VEGF-A quantification by ELISA in conditioned media (CM) from bCAFs after MCL-1 gene silencing (bCAFsgMCL-1) or not (bCAFsgCTRL). The bCAFs CM were generated during 72 h in EGM2 (Endothelial Cell Growth Medium-2) medium supplemented with 1% of FBS ( n = 6). Student t -test, ** P < 0.01. C qRT-PCR of VEGF-A, FGF2, and ANGPT1 mRNA in bCAFs treated or not by S63845 500 nM for 18 h normalized on RPLPO mRNA expression. Mean and SEM of three independent experiments are represented as relative quantity of mRNA. Student t -test, ** P < 0.01, ns non-significant. D VEGF-A quantification by ELISA in CM of bCAFs treated or not by S63845 500 nM for 18 h. Results were expressed as concentration (pg/ml) for 200,000 cells ( n = 5). Student t -test, ** P < 0.01. E Negative correlation (Spearman correlation coefficient r = −0.7381; p value = 0.0458) between VEGF-A concentration level in bCAFs CM and MCL-1 protein level (relative to actin level) determined by western-blot analysis in 8 primary cultures of bCAFs between passage 2 and 4.

Article Snippet: Endothelial cells (ECs) were routinely cultured in Endothelial basal medium (EGM2; containing 2% fetal bovine serum (FBS); PromoCell, Heidelberg, Germany) supplemented with Endothelial growth medium supplement pack (ECGM-2; PromoCell, Heidelberg, Germany) and 100 IU/mL penicillin and 100 mg/mL streptomycin on 0,1% gelatin-coated flasks.

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Generated, Concentration Assay, Western Blot

Journal: Cell Death & Disease

Article Title: MCL-1 as a molecular switch between myofibroblastic and pro-angiogenic features of breast cancer-associated fibroblasts

doi: 10.1038/s41419-025-07920-6

Figure Lengend Snippet: A bCAFs were treated for 18 h with MCL-1 inhibitor ( S63845 500 nM) or not. The treatment was removed and cells were rinsed and cultured for additional 72 h in EGM2 (Endothelial Cell Growth Medium-2) medium supplemented with 1% of FBS. After 72 h, the conditioned media (CM) were applied on HUVECs for tubulogenesis assay. B , C (Top) Representative images of tubulogenesis assay of HUVECs under ( B ) CM from bCAFs treated or not with MCL-1 inhibitor ( S63845 500 nM) or ( C ) CM from bCAFs expressing MCL-1 or not. (Bottom) Quantification of meshes, master junctions, and master segments of endothelial cells under ( B ) CM from bCAFs (untreated and S63845 ) n = 4 or ( C ) CM from bCAFs sgCTRL or sgMCL-1 ( n = 5). Student t -test, * P < 0.5, ** P < 0.01. D Chronological timeline of CAM model experimentation. After 10 days of embryonic development (ED10), T47D luminal breast cancer cell line with bCAFs expressing or not MCL-1 (bCAFsgCTRL or bCAFsgMCL-1) were xenografted. The tumours were treated with VEGF inhibitor (bevacizumab, BVZ 100 µg/CAM) every 2 days during one week until day 17 of embryonic development (ED17). E (Left) Representative pictures of engrafted tumours on CAM at ED17 are shown (Top: untreated or Bottom: treated with bevacizumab, BVZ 100 µg/CAM). (Right) Quantification of blood vessels density around the tumours (within a 5 mm radius of the tumour). Two-way ANOVA, *** P < 0.001, ** P < 0.01, ns non-significant.

Article Snippet: Endothelial cells (ECs) were routinely cultured in Endothelial basal medium (EGM2; containing 2% fetal bovine serum (FBS); PromoCell, Heidelberg, Germany) supplemented with Endothelial growth medium supplement pack (ECGM-2; PromoCell, Heidelberg, Germany) and 100 IU/mL penicillin and 100 mg/mL streptomycin on 0,1% gelatin-coated flasks.

Techniques: Cell Culture, Expressing