Figure 5

Colocalization of autophagosomes and lysosomes analysed by immunofluorescence. A Immunofluorescence of 3D4/21 cells that were infected with Ad-mRFP-GFP-LC3 and mock-infected, cultured in EBSS, treated with HCQ, or infected with AH at an MOI of 200 after 6 h. Thirty-two cells were randomly selected from each group; the yellow and red dots were counted for each cell. B Immunofluorescence of uninfected and infected PAMs collected from pigs and cultured in 24-well plates for 9 h. Yellow fluorescent puncta indicate the overlap of Mhp366 (green) and LAMP2 (red). Twenty cells were randomly selected from each group, and the yellow fluorescent puncta were counted from each cell. C Immunofluorescence of 3D4/21 cells cultured in EBSS medium or infected with AH after 6 h. Forty cells were randomly selected from each group, and the yellow fluorescent puncta were counted for each cell. Between-group differences were assessed using a t test; **p ≤ 0.01.