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. 2021 Feb 19;129(2):027005. doi: 10.1289/EHP7297

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EHP is an open-access journal published with support from the National Institute of Environmental Health Sciences, National Institutes of Health. All content is public domain unless otherwise noted.

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Figures 5A, 5B, 5C, 5D, 5E, 5F, 5G, and 5H are a set of internalization images of M D A-M D-231 cells, and Figure 5I is a bar graph. In Figures 5A, 5B, 5C, 5D, 5E, 5F, 5G, and 5H, images of M D A-M D-231 cells showing the effects of bisphenol S on epidermal growth factor with 50 micrometers are presented. Epidermal growth factor is exposed to 100 nanograms per alexa fluor 647-labeled epidermal growth factor (Figure 5A,E); bisphenol S plus epidermal growth factor is exposed to 1 microgram per milliliter bisphenol S plus 100 nanograms per milliliter epidermal growth factor (Figure 5B,F); bisphenol S 10 plus epidermal growth factor is exposed to 10 micrograms per milliliter bisphenol S plus 100 nanograms per milliliter epidermal growth factor, (Figure 5C,G);and epidermal growth factor plus epidermal growth factor is exposed to 100 nanograms per milliliter non-labeled epidermal growth factor plus 100 nanograms per milliliter epidermal growth factor (Figure 5D,H). The cells were exposed for 5 minutes. In Figure 5I, the bar graph titled M D A-M D-231 plots relative epidermal growth factor-label abundance, ranging from 0 to 120 in increments of 20 (y-axis) across epidermal growth factor, bisphenol S 1 plus epidermal growth factor, bisphenol S 10 plus epidermal growth factor, and epidermal growth factor plus epidermal growth factor (x-axis). — Effect of BPS on EGF internalization in MDA-MD-231 cells. (A–D) Representative images of EGF in MDA-MD-231 cells, (E-H) their respective post-processed images, and (I) quantification ( $mean \pm SEM$ ) following exposure to EGF* ( $100 ng$ /Alexa Fluor 647-labeled EGF) (A,E); $BPS 1 + EGF$ * ( $1 μ g / mL$ $BPS + 100 ng / mL$ EGF*) (B,F); $BPS 10 + EGF$ * ( $10 μ g / mL$ $BPS + 100 ng / mL$ EGF*) (C,G); and $EGF + EGF$ * ( $100 ng / mL$ nonlabeled $EGF + 100 ng / mL$ EGF*) (D,H). Images were taken 1 h after a 5-min exposure. The results were normalized by the number of cells and are expressed as the percentage of the EGF* group. $n = 4$ replicates/group. A generalized linear model was used to compare treatments. Different letters denote statistical differences among treatment groups at $p < 0.05$ . Colors in the processed images indicate the individual fluorescent signals detected by the software used to quantify the EGF internalization. Note: BPS, bisphenol S; EGF, epidermal growth factor; SEM, standard error of the mean.