Fig 1: Immunocytochemical analysis of HER2, MMP‐2, HIF‐1α, and COX‐2 expression in MCF‐7 breast cancer cells treated with IH, AS, and RO extract. Cells were stained with specific primary antibodies followed by Alexa Fluor–conjugated secondary antibodies, and nuclei were counterstained with Hoechst. (A, B, C, and E) Representative confocal ICC images of HER2 (A). MMP‐2 (B), HIF1a (C), and COX‐2 (E) expression under various treatment conditions. Scale bar = 10 μm; magnification 20×. (B) Quantitative analysis of HER2, MMP‐2, HIF‐1α, and COX‐2 expression based on immunocytochemistry (ICC). Fluorescence intensities were background‐corrected (Alexa Fluor 488 and 647 signal in untreated cells subtracted) and normalized to the control. Data represent mean ± SD (n = 3 independent experiments; 8 fields per condition per experiment); one‐way ANOVA with Tukey's multiple comparisons test.
Fig 2: Western blot analysis of the HER2/MMP‐2/COX‐2/HIF‐1α axis in MDA‐MB‐231 and MCF‐7 cells. (A, C) Representative Western blot bands for HIF‐1α, COX‐2, and MMP‐2 under CoCl₂ (D) or TNF‐α (E) stimulation, with and without plant extract treatment. (B, D) Quantitative densitometric graphs of HIF‐1α, COX‐2, MMP‐2, and HER2 expression levels following CoCl₂ or TNF‐α stimulation, with and without plant extract treatment. Protein levels were normalized to β‐Actin. Data represent mean ± SD (n = 3 independent experiments); one‐way ANOVA with Tukey's post hoc test.
Fig 3: Suppression of the MMP‐2 and COX‐2 in the TNF‐α‐induced models in MDA‐MB‐231 cells. (A, C) Representative confocal ICC images of MMP‐2 (G) and COX‐2 (H) expression under various treatment conditions. Scale bar = 20 μm; magnification 20×. (B, D) Quantitative analysis of MMP‐2 and COX‐2 expression based on immunocytochemistry (ICC). Fluorescence intensities were background‐corrected (Alexa Fluor 488 signal in untreated cells subtracted) and normalized to the control. Data represent mean ± SD (n = 3 independent experiments; 8 fields per condition per experiment); one‐way ANOVA with Tukey's multiple comparisons test.
Fig 4: The quantitative changes of TNF‐α (A), COX‐2 (B), VEGF‐α (C), MMP‐2 (D), and IL‐2 (E) in the bone marrow. Each independent experiment includes 5 rats with three repetitions per sample. C (control), DMBA (dimethylbenz(a)anthracene, cancer group), DIH (DMBA+Ínula helénium), DIHNN (DMBA+Ínula helenium+nor‐NOHA), DAS (DMBA+Alchemilla smirnovii Juz.), DASLN (DMBA+Alchemilla smirnovii Juz.+L‐NAME), DFU (DMBA+5‐fluorouracil), DFURO (DMBA+5‐fluorouracil+ Rumex obtusifolius ).
Supplier Page from Abcam for Rat MMP-2 ELISA Kit