Journal: Scientific Reports
Article Title: Stromal transcriptomics uncover LIF as a key effector in high tumor budding triple-negative breast cancer
doi: 10.1038/s41598-025-28439-y
Figure Lengend Snippet: The effects of LIF on TNBC cells and its proposed mechanism in cancer progression. a Representative image of migrated TNBC cells including MDA-MB-231 and MDA-MB-468 treated with a recombinant LIF compared to untreated condition as a negative control and IL-6 treatment condition as a positive control. b Bar graph shows the cumulative results of the number of migrated MDA-MB-231 and MDA-MB-468 cells from three independent experiments. c The PD-L1 expression in LIF-treated TNBC cells was evaluated by flow cytometry and the cumulative results from three independent experiments show PD-L1 in LIF-treated TNBC cells compared to the untreated condition. d Representative images of PD-L1 expression in LIF with or without the various doses of EC359 treatments in TNBC cells using flow cytometry. e Bar graph shows the cumulative results from three independent experiments. f Representative fluorescence images for untreated, LIF and LIF combined with EC359 treatments stained with anti-PD-L1 (green) antibody and Hoechst (blue). g A schematic diagram indicates several substances from cancer cells acting on CAFs in TNBC. Several signaling pathways in particular MAPK, ERK1/ERK2, and protein kinase B upregulated in high TB-CAFs which may lead to induce PD-L1 expression and increase cell migration in LIF-stimulated TNBC cells which could be attenuated by EC359. The *, **, and *** indicate significant significance of p < 0.05, 0.01, and 0.001, respectively. ns: not significance.
Article Snippet: The 2.5 × 10 4 of MDA-MB-231 and MDA-MB-468 TNBC cells which obtained from ATCC (Manassas, VA) were seeded in the upper chamber in 200 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\:\mu\:$$\end{document} L serum-free DMEM (Gibco; Thermo Fisher Scientific), and 600 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\:\mu\:$$\end{document} L of DMEM with 25 or 50 ng/ml recombinant LIF (R&D System Inc.) was added in the lower chamber as a chemoattractant compared to 1% FBS/DMEM as untreated condition and DMEM in presence of 50 ng/ml IL-6 (R&D system) as a positive control.
Techniques: Recombinant, Negative Control, Positive Control, Expressing, Flow Cytometry, Fluorescence, Staining, Protein-Protein interactions, Migration
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