Figure 7.

SNAPIN silencing in macrophages did not affect lysosome-autophagosome fusion. (A) Human macrophages were transfected with NS or SNAPIN siRNAs, incubated with 2 μM of rapamycin or 100 nM bafilomycin A₁ for 2 h. Cells were fixed with methanol for 20 min and stained with antibodies of LC3 (red) and LAMP1 (green). The highlighted areas in each panel were enlarged and shown in the inserted panels as indicated by the arrows. (B) Total LC3-positive puncta (red bars) and LC3 and LAMP1 colocalized puncta (yellow bars) were counted (numbers indicated in the bars) in 2 independent experiments. (C) The percentage of LC3⁺ LAMP1⁺ puncta to total LC3 puncta was calculated and presented. (D) Colocalization between LAMP1 and LC3 was measured by Pearson's correlation coefficient Rr employing NIS-element imaging software. *** represents p<0.001, following analysis by AVONA compare with NS-transfected cells. The presented images were processed by deconvolution.