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. 2023 Apr 21;13(4):707. doi: 10.3390/biom13040707

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© 2023 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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CAP treatment-induced cytotoxicity, apoptosis, and necrosis in MC38 cells in vitro via ROS/RNS generation. The MC38 cells are cultured for three days. Nine different exposure times and frequencies of CAP treatment are generated based on the different combinations of treatment frequencies and durations per treatment. (a,b) The normalized absorbance values of LDH (OD490) and CCK-8 (OD450) assays conducted with MC38 cells in both the control (mock) and PT groups, n = 4. (c,d) Flow cytometry analyses based on Annexin-V/PI staining to determine apoptotic and necrotic MC38 cells in the mock and PT groups (on-3, tw-3, and th-3), with logarithmic plots (c) and percentages of apoptotic cells (d), n = 3. (e,f) H₂O₂ and Nitrite concentrations in the MC38 cell culture supernatants in the mock and PT groups, n = 5. (g,h) Normalized fluorescence results for the levels of (g) intracellular ROS and (h) RNS in the MC38 cells treated with different CAP doses, n = 5. The data represent a minimum of three independent experiments (means ± SD). Statistical analysis is performed for all the PT groups using two-sided unpaired t-tests relative to the mock group (* p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001).

CAP treatment-induced cytotoxicity, apoptosis, and necrosis in MC38 cells in vitro via ROS/RNS generation. The MC38 cells are cultured for three days. Nine different exposure times and frequencies of CAP treatment are generated based on the different combinations of treatment frequencies and durations per treatment. (a,b) The normalized absorbance values of LDH (OD490) and CCK-8 (OD450) assays conducted with MC38 cells in both the control (mock) and PT groups, n = 4. (c,d) Flow cytometry analyses based on Annexin-V/PI staining to determine apoptotic and necrotic MC38 cells in the mock and PT groups (on-3, tw-3, and th-3), with logarithmic plots (c) and percentages of apoptotic cells (d), n = 3. (e,f) H₂O₂ and Nitrite concentrations in the MC38 cell culture supernatants in the mock and PT groups, n = 5. (g,h) Normalized fluorescence results for the levels of (g) intracellular ROS and (h) RNS in the MC38 cells treated with different CAP doses, n = 5. The data represent a minimum of three independent experiments (means ± SD). Statistical analysis is performed for all the PT groups using two-sided unpaired t-tests relative to the mock group (* p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001).