Shikonin Induces Glioma Necroptosis, Stemness Decline, and Impedes (Immuno)Proteasome Activity

Figure 3

Necroptosis inhibitors can significantly reduce Shikonin-induced U251 cell death. Images of Annexin V/PI doubling staining of U251 increasing concentrations of (a) necrostatin-1(O, 25, and 50 μM), (b) necrosulfomamide (O, 25, 50 μM), (c) GSK-872 (O, 10, and 20 μM), (d) Z-VAD-FMK (O, 10, and 20 μM) cells treated with Shikonin 12 μM for 24 hr and then subjected to flow cytometry. The ratios of apoptotic cells (Annexin V⁺/PI⁺ and Annexin V⁺/PI−) of (e) necrostatin-1, (f) necrosulfomamide, (g) GSK-872, and (h) Z-VAD-FMK were calculated. Data are expressed as mean ± SEMs (n = 5 per group). ()() versus untreated cells.

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