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. 2021 Aug 27;24(9):103038. doi: 10.1016/j.isci.2021.103038

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© 2021 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Flow cytometry and immunoblotting for apoptosis-related proteins

(A) Quantitative analysis of the expression levels of the anti-apoptotic protein Bcl-2. ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001 indicated significant differences compared to non-treated cells group. All the results are presented as mean ± SD; n = 3.

(B) Flow cytometric analysis of histograms of intracellular Bcl-2 protein at different times (2, 6, 12 hours) and at different radiation doses (0, 1, 2, 4 Gy).

(C) Quantitative analysis of the expression levels of the pro-apoptotic protein BAX. ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001 indicated significant differences compared to non-treated cells group. All the results are presented as mean ± SD; n = 3.

(D) Flow cytometric analysis of histograms of intracellular BAX protein at different times (2, 6, 12 hours) and at different radiation doses (0, 1, 2, 4 Gy).

(E) Quantitative analysis of P53 protein phosphorylation levels. ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001 indicated significant differences compared to non-treated cells group. All the results are presented as mean ± SD; n = 3.

(F) Flow cytometric analysis of intracellular P53 protein phosphorylation levels at different times (2, 6, 12 hours) and at different radiation doses (0, 1, 2, 4Gy).

(G) Quantitative analysis of the expression levels of the pro-apoptotic protein Cyto C. ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001 indicated significant differences compared to non-treated cells group. All the results are presented as mean ± SD; n = 3.

(H) Flow cytometric analysis of histograms of intracellular Cyto C protein at different times (2, 6, 12 hours) and at different radiation doses (0, 1, 2, 4 Gy).

(I) Western blot assay of the related apoptosis proteins after radiation treatment. HeLa cells were treated with different doses of irradiation (0, 1, 2, 4 Gy), and total proteins were extracted at different time points. The WB assay shows the changes of expression of apoptotic marker proteins (BAX, CytoC, Caspase-3, Cleaved Caspase-9, PARP) with time.