Fig. 2.

Celastrol treatment reduces number of cells containing Q57-YFP aggregates and increases Q57-YFP solubility. HeLa cells were transfected with Q57-YFP along with celastrol treatment at the concentrations indicated. a After 48 h of transfection, the formation of Q57-YFP aggregates was quantified using fluorescence microscopy. Visual fields which contained similar numbers of cells (based on the density of nuclei stained by Hoechst) were chosen under 20× objective, and then the number of aggregates in each field of vision was counted. Three different visual fields were quantified in each case, and data are shown as means ± SE (*P < 0.004, **P < 0.001, ***P < 0.0003, for each celastrol concentration treatment vs no celastrol). b To determine the amount of Q57-YFP monomer that could be solubilized from aggregates in lysates of the transfected cells by SDS treatment, the protein concentration of the insoluble fraction of the cell lysates was determined, and then 40 μg of protein was subjected to SDS solubilization treatment, followed by Western blot using anti-GFP antibody