Decreased lysosomal function.

<p>Effect of drug treatment on the abundance of p62 protein and mRNA (A); GAPDH protein and mRNA (B); phospholipid (C); and opsin (D). Quantification data in each panel is presented as fold change of total signal intensity compared to control (mean ± SD), * p ≤ 0.05. Tested drugs include chloroquine (CQ), fluoxetine (FX), imipramine (IP), dimebon (DM), tamoxifen (TM), chloropromazine (CP), amitriptyline (AT), and verapamil (VP).</p

TFEB, TFE3 and MITF promote lysosome biogenesis in response to lysosomotropic Drugs.

<p>Relative mRNA expression of cathepsin D and LAMP2 was measured using RT-PCR (A). Quantification data in each panel was presented as fold change of mRNA expression in the treatment group compared to control (mean ± SD), * p ≤ 0.05. Tested drugs include chloroquine (CQ), fluoxetine (FX), imipramine (IP), dimebon (DM), tamoxifen (TM), chloropromazine (CP), amitriptyline (AT), and verapamil (VP); Relative mRNA expression of cathepsin D, LAMP2, TFEB, TFE3 and MITF was measured using RT-PCR with TFEB, TFE3 and MITF knockdown and selected drugs (B), Quantification data is presented as fold change of mRNA expression either in the treatment group compared to DMSO control (mean ± SD) or under knockdown condition compared to control siRNA treatment (mean ± SD), * p ≤ 0.05.</p

Involvement of calcium in the nuclear translocation of TFEB, TFE3 and MITF, and lysosomal activation.

<p>Representative images for nuclear translocation of TFEB, TFE3 and MITF (A) and LTR staining (B) with and without BAPTA co-treatment. The graphs represent the percentage of total signal intensity with BAPTA co-treatment compared to corresponding drug treatment alone (mean ± SD), * p ≤ 0.05. Kinetic cytotoxicity profile with and without BAPTA co-treatment (C). Tested drugs include chloroquine (CQ), fluoxetine (FX), imipramine (IP), dimebon (DM), tamoxifen (TM), chloropromazine (CP), amitriptyline (AT), and verapamil (VP).</p

Increase of nuclear translocation of TFEB, TFE3 and MITF.

<p>Representative images for nuclear translocation of TFEB, TFE3 and MITF after 4 hrs (A) or 24 hrs (B) of drug treatment and for the time course of the nuclear translocation of TFEB (C), TFE3 (D) and MITF (E). Quantification data in each panel was presented as fold change of total signal intensity inside the nucleus compared to control (mean ± SD), * p ≤ 0.05. Tested drugs include chloroquine (CQ), fluoxetine (FX), imipramine (IP), dimebon (DM), tamoxifen (TM), chloropromazine (CP), amitriptyline (AT), and verapamil (VP).</p

Effect of lysosomotropic drugs on lysosome volume and pH.

<p>Representative images for LTR after 30 mins (A), 1 hr (B), 4 hrs (C) or 24 hrs (D) and lysoSensor green after 4 hrs (E) or 24 hrs (F) of drug treatment. Quantification data in each panel is presented as fold change of the total signal intensity compared to control (mean ± SD), * p ≤ 0.05. Tested drugs include chloroquine (CQ), fluoxetine (FX), imipramine (IP), dimebon (DM), tamoxifen (TM), chloropromazine (CP), amitriptyline (AT), and verapamil (VP).</p

Physicochemical properties, impact on autophagy and concentrations used in the studies.

<p>Physicochemical properties, impact on autophagy and concentrations used in the studies.</p

Increase of cathepsin B and D activity, and LAMP2 staining.

<p>Representative images for Cathepsin B activity (magic red staining) after 4 hrs (A) or 24 hrs (B), Cathepsin D activity (Bodipy-FL-P) after 4 hrs (C) or 24 hrs (D) and LAMP2 staining after 4 hrs (E) or 24 hrs (F) of drug treatment. Quantification data in each panel was presented as fold change of the total signal intensity compared to control (mean ± SD), * p ≤ 0.05. Tested drugs include chloroquine (CQ), fluoxetine (FX), imipramine (IP), dimebon (DM), tamoxifen (TM), chloropromazine (CP), amitriptyline (AT), and verapamil (VP).</p