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Fig. 2 | Journal of Translational Medicine

Fig. 2

From: Resibufogenin suppresses colorectal cancer growth and metastasis through RIP3-mediated necroptosis

Fig. 2

Resibufogenin induces necrosis in CRC cells. a Necrosis of colon cancer cells treated with PTL detected by flow cytometry at 24 h (n = 3 per group). See also Additional file 1 and Additional file 2: Figure S5. b Clone formation inhibition by resibufogenin from dose 0 to 1 μM in HCT116 cells. The panels represent the number of colonies as indicated. See also Additional file 1 and Additional file 2: Figure S2. c Flow cytometric analysis of resibufogenin treated cells with PE-annexin V/7-amino-actinomycin D staining (n = 3). Data derived from three separate experiments are presented as the mean ± SEM. See also Additional file 1 and Additional file 2: Figure S3. d Immunofluorescence analysis of PI-positive cells treated with resibufogenin. Cells were stained with Hoechst33342/PI (5 μg/ml) and subjected to analysis of necrosis population (n = 3). See also Additional file 1 and Additional file 2: Figure S4. e Cells were treated with the combination of resibufogenin and Z-VAD-fmk (20 μM). Cell viability was assayed were quantitatively analyzed (n = 6 per group). See also Additional file 1 and Additional file 2: Figure S5. f, g Morphological changes of resibufogenin-treated cells and tissues as observed by TEM. ×10,000 for all. Scale bar = 1 μm. h The resibufogenin-induced LDH leakage was increased in a dose-dependent manner (n = 3). i Effects of resibufogenin on the ROS generation of HCT116 cells (n = 3). See also Additional file 1 and Additional file 2: Figure S6. j The effect of resibufogenin on cell viability of colon cancer cells treated with L-NAC (n = 6). Data presented are showed as mean ± SEM from three independent experiments. *P < 0.01 indicate significant difference

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