Figure 3. Glutamine consumption and GLS activity are important for optimal adenovirus replication.

(a) NHBE cells were infected with AD WT at an MOI of 1 in the presence (+ Q) or absence (−Q) of 2.5 mM glutamine. At 24 h post infection, progeny virus was harvested and infectious virus titer was measured using a TCID₅₀ end point dilution plaque assay. (b) (bottom) Glutamine consumption rates of NHBE cells stably expressing scrambled shRNA (shCTR, blue bars) or GLS shRNA (shGLS, red bars) 24 h post mock infection or infection with AD WT (bottom). (top) Immunoblotting depicting levels of GLS splice isoforms KGA and GAC in NHBE cells stably expressing scrambled shRNA (shCTR) or GLS shRNA (shGLS). (c) Progeny virus was collected from NHBE cells stably expressing shCTRL or shGLS 24 h post infection with AD WT, and adenovirus titres were determined as in (a). (d) NHBE cells were treated with 1.0 μM CB-839 or DMSO, and either mock-infected or infected with AD WT. Glutamine consumption rates were measured 24 h post treatment and infection. (e) Progeny virus was harvested from NHBE cells treated with DMSO or 1.0 μM CB-839 24 h post infection with AD WT, and adenovirus titers were determined as described in (a). For (a–e), error bars denote s.d. (n=3), **P<0.01. Student's t-test.