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. 2023 Aug 14;325(3):C708–C720. doi: 10.1152/ajpcell.00199.2023

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Licensed under Creative Commons Attribution CC-BY 4.0. Published by the American Physiological Society.

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Figure 4. — mRNA levels of epidermal growth factor receptor (EGFR; A) and human epidermal growth factor receptor 2 (HER2; B) in MCF-7, MCF-7 cells resistant to fulvestrant (MCF-7/Fulv), and MCF-7 cells resistant to tamoxifen (MCF-7/Tam) measured by RT-qPCR. Results are expressed as mean ± SD of relative mRNA level of three independent experiments. *Statistically significant differences (P ≤ 0.05) compared with parental MCF-7 cells. C: colocalization between members of EGFR/HER family receptors on MCF-7, MCF-7/Fulv, and MCF-7/Tam determined by proximity ligation assay. D: immunoblotting of pEGFR/EGFR, pAKT/AKT, pERK1/2/ERK1/2, and actin constitutively present in MCF-7, MCF-7/Fulv, and MCF-7/Tam cells. E: the ratio of phosphoEGFR (pEGFR) over EGFR, phosphoAKT (pAKT) over AKT, and phosphoERK1/2 (pERK1/2) over ERK1/2 was quantified from band density using ImageJ, and the values are displayed relative to the levels observed in the parental MCF-7 cells. Results are expressed as mean ± SD of % change of relative band density of three independent experiments. *Statistically significant differences (P ≤ 0.05) compared with parental MCF-7 cells. F: effect of epidermal growth factor (EGF) on cell proliferation in MCF-7, MCF-7 cells resistant to fulvestrant (MCF-7/Fulv), and MCF-7 cells resistant to tamoxifen (MCF-7/Tam) estimated by MTT assay. Cells were seeded in 48-well culture plates (10,000 cells/well) and cultured in RPMI without phenol red supplemented with 10% charcoal-stripped serum (CSS) in the presence or absence of EGF for 48 h. Results are expressed as mean ± SD of % change to control untreated cells of at least four independent experiments. *Statistically significant differences (P ≤ 0.05) compared with control untreated cells. MTT, 3-[4,5-dimethylthiazol-2-yl]-2,5-dimethyltetrazolium bromide.