Figure 3
Treatment with UA-8 modulates the autophagic response in HL-1 cells during starvation. (a) Formation of LC3-II protein in starved HL-1 cells. Left panel: representative western blots demonstrating the time course accumulation of LC3-II in starved cells. Right panel shows the results of western blot quantification after 2 and 24 h of starvation, respectively. (b) Representative images following 24 h of starvation in HL-1 cells immunostained to detect LC3 positive puncta (green), a marker of autophagy. Nonstarved HL-1 cells were treated with chloroquine (50 μM), a blocker of autophagosomal degradation, as a control. Images were acquired with a Zeiss Axio Observer epifluorescence microscope using a × 63 objective (Oberkochen, Germany). Alexa Fluor 488 was conjugated LC3 Ab (green) and DAPI nuclear stain (blue) were utilized. (c) Representative electron micrograph (EM) images of nonstarved HL-1 cells and cells starved for 24 h with and without UA-8. White arrows identify autophagosomal vacuoles; note mitochondrial engulfment. Values are represented as mean±S.E.M., N=3. Significance was P<0.05, *significantly different from control nonstarvation, #significantly different from UA-8
