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. 2021 Oct 22;12:6144. doi: 10.1038/s41467-021-26423-4

Fig. 8. Reduced aspartate levels rescue the survival of Ripk1−/− cells from starvation.

Fig. 8

a, b Cell survival of Ripk1−/− MEFs responding to AOA treatment was assessed by trypan blue exclusion. WT and Ripk1−/− MEFs were cultured in EBSS for 6 h with 0.5 mM or 0.75 mM AOA treatment as indicated. Quantification of cell survival under different conditions was showed in (b). Data was expressed as the rate of viable cell number to dead cell number. Bars represent mean ± SEM (n = 9, three biologically independent samples per group, three images in each biological repetition). Scale bar represents 10 μm. P values were determined by one-way ANOVA by Tukey’s multiple comparisons test. c Intracellular aspartate levels in RIPK1−/− HEK293T cells transfected with pcDNA-3.1 or pcDNA-3.1-SP1 plasmids were detected by LC–MS. Bars represent mean ± SEM (n = 6 biologically independent samples per group). P values were determined by a two-tailed Student’s t-test. d, e Cell survival of RIPK1−/− HEK293T cells responding to SP1 overexpression was assessed by trypan blue exclusion. Cells were transfected with pcDNA-3.1 or pcDNA-3.1-SP1 plasmids. The medium was changed to culture medium or EBSS for 6 h before harvest. Quantification of cell survival under different conditions was showed in (d). Data was expressed as the rate of viable cell number to dead cell number. Bars represent mean ± SEM (n = 9; three biologically independent samples per group; three images in each biological repetition). Scale bar represents 10 μm. P values were determined by a two-tailed Student’s t-test. f Graphical summary illustrating the role of RIPK1 in starvation resistance.