Figure 4
From: AMPK contributes to autophagosome maturation and lysosomal fusion
AMPK α1 knockout interferes with the formation of autolysosomes. (A) Control cells and AMPK α1 knockout cells were transfected with the plasmid encoding mRFP-LC3 and treated with compound C for 18 h. Cells were fixed and stained with an anti-LAMP1 antibody. Bars: 10 μm. (B) Ratio quantification of autophagosomal LC3 puncta (RFP-LC3 only) to autolysosomal LC3 puncta (RFP-LC3 and LAMP-1) Each data represents the mean and standard deviation of two independent experiments (n = 30), control cells vs AMPK Knockout cells, *P < 0.05. (C) Representative electron micrograph of HEK293T cells and AMPK α1 knockout cells after compound C treatment. Yellow arrows denote the autophagosome; blue arrows, autolysosomes; and red arrows, lysosomes. Bars: 1 μm. (D) Ratio quantification of autophagosomal puncta to autolysosomal puncta. We counted autophagosomes and autolysosomes in 54 individual cells (n = 54) of each group, and calculated the relative abundance. Graph shows the average and standard deviations. Control cells vs. knockout cells, *P < 0.0001.
