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. Author manuscript; available in PMC: 2021 May 17.
Published in final edited form as: Cell Rep. 2021 May 4;35(5):109076. doi: 10.1016/j.celrep.2021.109076

Figure 5. SphK2/S1P signaling via HDAC1/2 inhibition induces CerS6/ceramide-dependent mitophagy in aging T cells.

Figure 5.

(A and B) Expression of CerS6 mRNA in the absence (A) or presence of HDAC inhibitor MS275 (B) was measured by qPCR in ex vivo TCR activated T cells isolated from Y and A WT, SphK2−/− (SK2), or CerS6−/− mice. Data are means ± SDs from 3 independent experiments (n = 3).

(C and D) Activated T cells obtained from Y and/or A mice were cultured with/without MS275, and mitophagy/autophagy was detected by live-cell imaging using MTR-LTG colocalization (arrows indicate colocalized signals), and scale bars indicate 1 μm. Images represent at least 3 independent experiments. Quantification of colocalization (Rc) (C, lower panel). LC3 activation as measured by Cyto-ID (D). Data are means ± SDs from 3 independent experiments (n = 3). *p < 0.05 as determined by Student’s t test.

(E) TCR-activated T cells in the presence of splenocytes isolated from Y Pmel mice were co-cultured with B16 melanoma cells for 1, 6, and 16 h in the absence/presence of MS275. Data are means ± SDs from at least 3 experiments (n = 3). *p < 0.05 as determined by Student’s t test.