Figure 2.

Autophagy-Independent Recruitment of LC3-I onto MHV-Induced DMVs

(A and B) HEK293 cells stably transfected (B) or not (A) with a plasmid expressing GFP-LC3 were infected with MHV-Srec and processed for IF at 7 hr p.i.

(C) Summary statistics of the samples shown in (A) and (B) expressed as the percentage of LC3 or GFP-LC3 puncta colocalizing with the nsp2/nsp3 signals. Error bars represent the standard error of the mean percentage from counting 40 cells in three independent experiments. The asterisk indicates that the two samples are significantly different (t_{df = 78} = 10.4; p < 0.00001).

(D) Atg7^+/+ and Atg7^−/− MEF were infected with MHV before being processed for IF at 7 hr p.i.

(E) Statistical analysis of the samples shown in (D) performed as described in (C).

(F) HeLa-CEACAM1a cells were infected with MHV-nsp2GFP for 7 hr before fractionating a cell extract on a continuous Optiprep gradient. Ten fractions were collected from the top to the bottom of the gradient and probed with antibodies against EDEM1, GFP, and LC3. The fractionation profile was confirmed by performing this experiment three times.

(G) HeLa-CEACAM1a cells were transiently transfected with a plasmid expressing C-terminally HA-tagged, nonlipidable LC3 before being infected with MHV. Cells were fixed at 7 hr p.i. and processed for IF. DMVs and LC3-HA were detected with antibodies against nps2/nsp3 and HA, respectively.

See also Figure S1.