Reactive Oxygen Species Regulate Endoplasmic Reticulum Stress and ER-Mitochondrial Ca²⁺ Crosstalk to Promote Programmed Necrosis of Rat Nucleus Pulposus Cells under Compression

Figure 2

Ca²⁺ assembling in mitochondria may contribute to ERS-related Ca²⁺ release. (a, b) Representative histograms and statistical analysis of [Ca²⁺]_m detected by flow cytometry in compression-treated NP cells with Rhod-2 AM. NS means no significant difference. The values are expressed as the from three biological replicates ( and vs. control, ANOVA/LSD). (c) Typical fluorescence photomicrograph of in situ [Ca²⁺]_m staining with Rhod-2 AM. (d, e) Representative histograms and statistical analysis of [Ca²⁺]_m detected by flow cytometry in compression-treated NP cells with Rhod-2 AM. Cells were pretreated with 200 μM 4-PBA for 1 h and then subjected to compression for 36 h. The values are expressed as the from three biological replicates ( vs. control, ^## vs. compression alone, ANOVA/LSD). (f) Typical fluorescence photomicrograph of in situ [Ca²⁺]_m staining with Rhod-2 AM. Cells were treated as in (d). (g, h) Representative histograms and statistical analysis of [Ca²⁺]_m detected by flow cytometry in compression-treated NP cells with Rhod-2 AM. Cells were pretreated with 50 μM DAN or 2.5 μM XeC for 1 h and then subjected to compression for 36 h. NS means no significant difference. The values are expressed as the from three biological replicates ( vs. control, ^## vs. compression alone, ANOVA/LSD). (i) Typical fluorescence photomicrograph of in situ [Ca²⁺]_m staining with Rhod-2 AM. Cells were treated as in (g).

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