Figure 5.

Re-submersion Induces Cellular Stresses Mediated by Low Oxygen Tension

(A) Top upregulated pathways in monolayers 8 h after re-submersion as compared to ALId21.

(B) A schematic summary of the cellular pathways activated by low oxygen tension.

(C) Hif1α protein accumulation evaluated by immunoblot of cell lysates derived from monolayers re-submerged for 0, 2, 4, 8, and 24 h.

(D) Hif1α targets expression defined by qPCR assays in monolayers re-submerged for 0, 2, 4, 8, 24 h and 7 days. Results were expressed relative to the 0 h time point. Mean ± SD are shown. n = 3 samples/time point.

(E) Intracellular lactate levels in monolayers as a function of time after re-submersion. n = 3 samples/group.

(F) Changes in levels of proteins known to be involved in the UPR pathway defined by immunoblotting of cell lysates as a function of time after re-submersion.

(G) Expression of UPR target genes, defined by qPCR, in monolayers at different times after re-submersion. Results were expressed relative to the 0 h time point. Mean ± SD were plotted. n = 3 samples/group.

Two-tailed Student’s t test relative to 0 h: ^∗p < 0.05; ^∗∗p < 0.01. Immunoblots are representative of two independent experiments.