Figure 2.

Morusin induces autophagy at early stages. A. Morusin induces autophagy at the initiation step. HeLa cells expressing GFP-RFP-LC3 were treated with morusin alone or in combination with 3-MA or chloroquine (CQ). Images of LC3 puncta containing green and red fluorescence were obtained with a confocal fluorescent microscope at an excitation wavelength of 488 nm and 543 nm, respectively. In the zoomed images, the arrows indicate red LC3 puncta with weak green fluorescence, and the arrowheads indicate red and green puncta colocalization. B. The number of LC3 puncta was counted in three different arbitrary areas, and is shown on the graph. Data are presented as the mean ± SEM. Morusin and CQ additively induce LC3-II accumulation (left panel). The relative ratio of yellow and red puncta is presented (right panel). C. The levels of LC3-II in cells treated with both morusin and CQ was compared to levels in cells treated with either drug alone.