Journal: Journal of Translational Medicine
Article Title: NGF-mediated tumor–stroma crosstalk promotes prostate cancer aggressiveness
doi: 10.1186/s12967-026-07915-7
Figure Lengend Snippet: PC-derived CAFs release NGF and express mesenchymal/myofibroblast markers as well as TrkA. ( A ) Quantification of NGF secretion by ELISA in conditioned media collected from primary CAFs lines (C1–C8) isolated from independent PC patients at Gleason’s scores 6–8. Data are expressed as pg/ml (mean ± SD) from three independent experiments performed in duplicate. ( B ) Representative IF images of NGF expression in primary CAFs (C1). NGF is shown in green and nuclei in blue. Images are representative of three independent experiments. Scale bar, 10 µM. ( C ) Quantitative analysis of NGF fluorescence intensity in primary CAF cultures (C1–C8). Mean fluorescence intensity (MFI) was normalized to the number of nuclei and expressed as fold increase relative to the fluorescence from the secondary antibody, used as control (II Ab). Data are presented as mean ± SD from several independent fields analyzed for each culture. ( D ) Western blot analysis of TrkA and mesenchymal/myofibroblast markers in lysates from primary prostate CAFs (C1–C8). Vimentin was used as a mesenchymal marker, while α-SMA and FAP-1 were used as myofibroblast-associated markers. Tubulin served as loading control. LNCaP cells, positive for the expression of E-cadherin, were included as epithelial control
Article Snippet: The following primary antibodies were used: mouse monoclonal antibodies against α-tubulin (#E-AB-20036, Elabsciences, Houston, TX, USA), GAPDH (#E-AB-20079, Elabsciences), α-SMA (CGA7, Santa Cruz Biotechnology, Dallas, TX, USA), FAP (SS-13, Santa Cruz Biotechnology), and E-cadherin (Clone 36, RUO); rabbit polyclonal antibodies against vimentin (H-84, Santa Cruz Biotechnology, Dallas, TX, USA) and TrkA (06-574, Millipore, Burlington, MA, USA).
Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Isolation, Expressing, Fluorescence, Control, Western Blot, Marker