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. 2017 Mar 29;8(19):31923–31937. doi: 10.18632/oncotarget.16652

Figure 5. Inhibition of autophagy enhances Fhit-induced cell death.

Figure 5

(A) Effects of autophagy inhibition on viability of H460 cells transduced with Ad-Fhit. H460 cells, stably expressing shRNA against beclin-1 (shBeclin-1) or LC3B (shLC3) to block autophagy, were transduced with Ad-Fhit and cell viability was analyzed by a WST assay. Results are mean percentage ± standard deviation of three independent experiments. ††p < 0.01 and †††p < 0.001 for the Fhit transduction with beclin-1 or LC3 shRNA treatment versus the Fhit transduction with control shRNA (shControl) treatment. MOI, multiplicity of infection. (B) Western blot analysis of apoptotic protein expression after Fhit transduction in H460 cells stably expressing shRNA against beclin-1 (shBeclin-1) or LC3B (shLC3). shControl, control shRNA. (C) Effects of 14-3-3τ inhibition on viability of H460 cells transduced with Ad-Fhit. H460 cells were transfected with siRNA against 14-3-3τ (si14-3-3τ) and then transduced with Ad-Fhit. Cell viability was analyzed by a WST assay. Scrambled siRNA (siControl) was used as a nonspecific siRNA control. Results are mean percentage ± standard deviation of three independent experiments. †††p < 0.001 for the Fhit transduction with 14-3-3τ siRNA treatment versus the Fhit transduction with control siRNA treatment. MOI, multiplicity of infection. (D) Western blot analysis of apoptotic protein expression after Fhit transduction in H460 cells that had been transfected with siRNA against 14-3-3τ (si14-3-3τ). siControl, control siRNA. (E) Effects of pharmacological inhibition of autophagy by hydroxychloroquine (HCQ) on the viability of H460 cells transduced with Ad-Fhit. H460 cells were transduced with Ad-Fhit and then treated with 10 μM of HCQ or PBS (vehicle) as a control. Cell viability was assessed by a WST assay. Results are presented as mean percentage ± standard deviation of three independent experiments. †††p < 0.001 for the Fhit transduction with HCQ treatment versus the Fhit transduction with vehicle treatment. (F) Western blot analysis of apoptotic protein expression in H460 cells that were transduced with Ad-Fhit and treated with 10 μM of HCQ.