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. 2013 Nov 22;8(11):e82481. doi: 10.1371/journal.pone.0082481

Figure 4. SQSTM1/p62 abundance assessment.

Figure 4

U2OS cells stably expressing GFP-LC3 were incubated with compounds for 24 h. SQSTM1/p62 protein level was evaluated using an ELISA assay described in Materials and Methods. The fold change of SQSTM1/p62 abundance compared to DMSO control was plotted for each compound and the data are expressed as mean ± SD. The dotted line depicts the DMSO control level. Rapamycin (1 µM) and bafilomycin A (5 nM) were tested at single concentration. Based on viability profile three concentrations were chosen for the compounds. The concentration range tested is from low (left) to high (right). For chloroquine, desipramine, imipramine concentrations of 15 µM, 30 µM and 50 µM were selected. For clomipramine, fluoxetine, nortriptyline, paroxetine concentrations of 10 µM, 15 µM and 20 µM were selected. For astemizole concentrations of 5 µM, 10 µM and 25 µM were selected. For chlorpromazine, sertraline and thioridazine concentrations of 2.5 µM, 5 µM and 10 µM were selected. All concentrations have viability greater than 60% except the ones labeled with “†”.