Fig 2: Inhibition of dynein activity decreased APLs formation. Mouse CAMs were stained with acridine orange for 17 min. Representative dot plots of flow cytometry (A) and summarized red-to-green fluorescence ratio analysis showing APLs formation in CAMs with DCTN2 cDNA (B) or EHNA (C). (D) Summarized red-to-green fluorescence ratio analysis showing APLs formation in CAMs with spautin-1 (10 μM) and leupeptin (0.25 mM). (E) Representative Western blot documents showing the expression of Lamp-1 from CAMs. (F) CAMs were treated with chloroquine (CQ, 100 μM) for 30 min. or left untreated. They were then stained with LysoSensor Green DND-189 and analysed by fluorescence microscopy (n = 6 for all panels). *P < 0.05 versus Scramble; #P < 0.05 versus CAMs with X/XO or H2O2 or Nox1 cDNA transfection alone.

Fig 3: Dynein inhibition increased accumulation of AP and decreased breakdown of AP. (A and B) Summarized per cent of Cyto-ID-stained cells showing the relative number of AP in CAMs with EHNA or DCTN2 cDNA. (C and D) Representative confocal images and summarized co-localization coefficient showing the co-localization of ubiquitin with p62 (n = 6 for all panels). *P < 0.05 versus Ctrl or Scramble; #P < 0.05 versus CAMs treated with X/XO or H2O2 or Nox1 cDNA transfection alone.

Fig 4: Inhibition of dynein activity blocked lysosome fusion. (A) Representative confocal microscopic images showing the co-localization of LC3B-GFP with Lamp1-RFP in live CAMs. (B) Summarized co-localization coefficient (PCC) of LC3B and Lamp1 in CAMs transfected with scramble or DCTN2 cDNA. (C) Summarized co-localization coefficient of LC3B-GFP and Lamp1-RFP in CAMs with or without EHNA (30 μM; n = 6 for all panels). *P < 0.05 versus Ctrl or Scramble; #P < 0.05 versus CAMs treated with X/XO or H2O2 or Nox1 cDNA transfection alone.