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. 2020 May 15;21(10):3491. doi: 10.3390/ijms21103491

Figure 4.

Figure 4

Effect of nonylphenol (NP) on Sertoli cells during mouse testicular fraction (MTF) culture. (A) Representative immunoblots of SOX9, a Sertoli cell-specific marker, in 30-day cultured MTFs with or without NP. β-actin was used as the loading control. (B) Ratios of SOX9 to β-actin are shown as the mean and the standard error of the mean (n = 5). No significant difference was noted between the 1, 10, and 50 µM NP-treated MTFs and controls. (C) Double immunohistochemistry of the MTF sections was used to examine the localization of SOX9 and vimentin in MTFs cultured for 30 days with NP and with the IgG isotype-negative control. Scale bars = 50 µm. All of the images were acquired at the same magnification. (D) The average number of SOX9-positive Sertoli cells in the tubules was calculated on the basis of immunostaining. At least 50 tubules were scored for each MTF (5–6 biological replicates). The expression of SOX9 and number of SOX9-positive cells in each tubule did not differ significantly between the NP-treated and untreated MTFs. The data are presented as mean ± standard error.