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. 2023 Jan 18;9:12. doi: 10.1038/s41420-023-01320-5

Fig. 3. SPHK2 is essential for LPS-triggered oxidative stress and NLRP3 inflammasome activation in macrophage.

Fig. 3

A sh-SPHK2 or shNC transfected RAW264.7 cells were stained with JC-1 and analyzed quantitation of fluorescence intensity by flow cytometry. B sh-SPHK2 or shNC transfected RAW264.7 cells were stained with MitoSOX and analyzed by flow cytometry. LPS-elicited mtROS generation was inhibited by SPHK2 knockdown. C qRT-PCR analysis was performed to determine the correlation between SPHK2 and NLRP3 mRNA levels for individual sample. D Western blots of NLRP3 in RAW264.7 cells exposed to PBS or LPS, treating with or without ABC294640. E Cell lysates were immunoblotted for NLRP3, ASC, and Caspase-1 p20 proteins in RAW264.7 cells. F Representative confocal microscopic images of gene-modified RAW264.7 cells co-localization with Caspase-1 p20 (red) and NLRP3 (green). bar = 100 μm. Data were presented as the means ± SEM from at least three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.005; N.S., not significant.