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. 2021 May 25;11(15):7337–7359. doi: 10.7150/thno.57804

Figure 4.

Figure 4

MRPL52 resists the caspase-dependent mitochondrial apoptosis of BC cells exposed to hypoxia. (A) Cellular apoptosis examined by TUNEL staining (mean ± SD, n = 3). *P < 0.05. Scale bars, 100 µm. (B) MMP detected by JC-1 probe. JC-1 red (JC-1 aggregate) shows healthy MMP, whereas JC-1 green (JC-1 monomer) shows decreased MMP. ΔΨm was represented as the red/green ratio (mean ± SD, n = 3). *P < 0.05. Scale bars, 100 µm. (C) Alterations in mitochondrial permeability transition pore opening were detected by calcein AM staining. The weaker the green fluorescence in cells, the higher the degree of opening of mPTP (mean ± SD, n = 3). *P < 0.05. Scale bars, 100 µm. The proteins expression of (D) Cytochrome c in cytoplasm and mitochondria, (E) cleaved PARP, cleaved Caspase-3, GPX4 and Caspase-1 were analyzed using WB. Cyto, cytosolic protein; Mito, mitochondrial protein. (F-G) MDA-MB-231 cells were incubated with Z-VAD-FMK (50 μM), a pan-caspase inhibitor or ferrostatin-1 (20 nM), a ferroptosis inhibitor or necrosulfonamide (20 nM), a necroptosis inhibitor for 24 h. (F) Cell viability, (G) cleaved PARP and cleaved Caspase-3 levels were measured using CCK-8 and WB analysis, respectively (mean ± SD, n = 3). *P < 0.05.