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. 2022 Feb 2;76:103847. doi: 10.1016/j.ebiom.2022.103847

Figure 4.

Fig 4

Gpx4 downregulation makes chondrocytes more sensitive to oxidative stress. (a) The efficacy of Gpx4 specific shRNA were measured by qRT-PCR and western blotting in MCC. (b) Cell viability and death rate of MCC-shNC and shGpx4 treated with different concentration TBHP for 24 h measure by CCK8. Data of TBHP 10μM, 15μM, 20μM, 30μM were relative to its corresponding cell line data in TBHP 0. (c) Cell viability of MCC-shNC and shGpx4 treated with different concentration TBHP and rescued via Fer-1, DFO and Nec-1, repectively. (d) MDA contents of MCC-shNC and shGpx4 and treated with TBHP 30μM for 4 h. Data of TBHP were relative to its corresponding cell line data of Ctrl. (e) Glutathione peroxidase activity level of MCC-shNC and shGpx4. (f) GSH contents of MCC-shNC and shGpx4 (left) and treated with TBHP 30μM for 4 h (right). Data of TBHP were relative to its corresponding cell line data of Ctrl. (g) Lipid ROS level was analyzed using Liperfluo fluorescence probe of MCC-shNC and shGpx4. Scale bar, 50 μm. GFP indicate lentivirus affected. (h) Intracellular Fe2+ level was detected using FerroOrange Assay of MCC-shNC and shGpx4. GFP indicate lentivirus affected. Scale bar, 75 μm. Data are expressed as mean ± SD, * P< 0.05; ** P< 0.01; *** P< 0.001. Student's t-test and one-way ANOVA were used for comparison between two groups and multiple groups, respectively.