Figure 4. Expression of WT RNase L promotes death of RNASEL-ADAR1 KO cells.

(a) WT and KO A549 cells as indicated were transfected with 20 ng/ml of pIC and assessed for dead cells and total cells. Four separate wells were treated with each experimental condition and a minimum of 4 image fields (>10000 cells per well) were analyzed per well. Data represent the means SD from a minimum of four independent replicates. Similar data were obtained from two additional independent experiments. (The data for WT cells are the same as shown in Figure 3b). (b) RNASEL KO or RNASEL-ADAR1 DKO cells transduced with lentiviruses expressing either WT RNase L or inactive mutant (R667A) RNase L, lysed and proteins analyzed by Western immunoblotting with anti-Flag M2 monoclonal antibody (upper) or with anti-β-actin antibody (lower). (c) RNASEL KO or RNASE L-ADAR1 DKO cells were transduced with lentiviruses expressing either WT RNase L or inactive mutant (R667A) RNase L and dead and total cells assessed in the IncuCyte system. A minimum of four separate wells was treated with each experimental condition and a minimum of 9 image fields were analyzed per well. Data represent the means SD and are from one of two representative experiments. Movies showing cells from 0–72 hr post transduction are shown in Videos 1–4. See Figure 4—source data 1.

DOI: http://dx.doi.org/10.7554/eLife.25687.011