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. 2021 Oct 20;11:755433. doi: 10.3389/fonc.2021.755433

Figure 2.

Figure 2

Extracellular release of HMGB1 and ATP following Cabozantinib treatment of DU-145 and PC-3 prostate cancer cell lines. (A) Western blot analysis of HMGB1 expression in cytosolic membrane fraction of untreated (NT) or Cabozantinib-treated (2.5 μg/ml) DU-145 cells. β-Tubulin was employed as internal reference standard. Bands intensity was measured by ImageJ software; histograms represent the ratio between HMGB1 (25–29 kDa) and β-tubulin (55 kDa) band intensities. (B) HMGB1 release upon Cabozantinib treatment. Histograms represent the ratio between the median values of HMGB1 released by treated cells (2.5 and 5 μg/ml) vs. values of HMGB1 released by untreated cells (NT). (C) Release of ATP induced by Cabozantinib. The ATP release was measured by ENLITEN-Promega KIT as luminescence signals. The histograms represent the mean value of ATP moles of two independent experiments in duplicate ± SEM. The line on histograms represents the NT (not treated) value. *p < 0.05; **p < 0.01; ***p < 0.001, Student’s t-test. NT, untreated cells as experimental control.