FIGURE 7.

Lacritin restores metabolism. A, HCE-T cells were stressed overnight with IFNG and then treated for different times with 10 nm lacritin or C-25 in the presence of TNF. The oxygen consumption rate was monitored in the absence or presence of oligomycin (500 nm), FCCP (375 nm), or antimycin A (750 nm) and rotenone (1 μm) together. B, features of the oxygen consumption rate as revealed by use of oligomycin, FCCP, antimycin A, and rotenone (t test; **, p < 0.01). C, HCE-T or HCE-T shATG7(1) cells were stressed overnight with IFNG and then stained for 30 min with MitoTracker Red FM. After addition of lacritin or C-25 (10 nm) in the presence of TNF, live mitochondria were monitored at 20-s intervals for 30 min, and fission frequency was estimated from ComponentCount of Mytoe. D, schematic diagram of mitochondrial fusion or fission. E, HCE-T cells were stressed overnight with IFNG and then treated for 10 min in replicates of six with 10 nm lacritin or C-25 or left untreated in the presence of TNF. Metabolites in cell lysates were identified by mass spectrometry. Values were analyzed by the t test and non-parametric Wilcoxon test of the log-transformed normalized data (*, p < 0.05) and further analyzed by hierarchical clustering. Error bars represent S.E. lacrt, lacritin; antimyc, antimycin A; rot, rotenone; res, respiration; res cap, respiratory capacity; untreat, untreated.