Figure 4

LC3 recruitment by selected CRPs in ZF4 cells and in zebrafish larvae. (A) Representative confocal images of the FITC immune-labelled LC3B in ZF4 cells treated with either GFP or the CRP-mix for 4 h. Nuclei were stained with DAPI. Autophagosome levels were quantified as the area (per cell) of over-threshold green fluorescence corresponding to the intracellular puncta and represented as fold changes in comparison to the GFP treatment as determined by the following formula: over-threshold fluorescence per cell in CRP-mix-treated monolayers/over-threshold fluorescence per cell in GFP-treated monolayers. This experiment was performed 3 times, each in triplicate. Symbol’a’ indicates statistically significant differences between CRP-mix and GFP treatments at the P < 0.05 level. Data were analysed by using two-tailed unpaired Student’s t-test. (B) Representative images of GFP-LC3 transgenic zebrafish larvae at 3 days post injection with 150 pg of pMCV1.4 or pMCV1.4-crp1/crp4/crp5/il6 plasmid constructs. Corresponding scale bars equal 50 and 100 µm.