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. 2010 Dec 23;5(12):e14410. doi: 10.1371/journal.pone.0014410

Figure 4. Niclosamide affects the formation of the ubiquitin-enriched aggregates caused by proteasome inhibition in SH-SY5Y cells.

Figure 4

(A) Signal intensities of the aggregates in SH-SY5Y cells stably expressing GFP-ubiquitin treated for 8 h with different autophagy inducers. The following concentrations were used: niclosamide (1, 3 and 10 µM), rottlerin (0.1, 1, 3 µM) and perhexiline (1, 3, 10 µM) alone (light grey), and together with 5 µM MG132 (dark grey). Mean intensities (with standard deviations) were measured in three independent wells for each condition using the automated high-content fluorescence imager. The p values for the designated samples calculated by Student's t-test were 0.03 and 0.0014, respectively. (B) Representative images of cells incubated as indicated with 5 µM MG132 and 10 µM niclosamide for 8 h prior to PFA fixation. Scale bar represents 10 µm. Preceding time points are shown in Fig. S3C. (C, D) Same as in A, using 1 µM epoxomycin, 5 µM MG132, 10 µM niclosamide or 10 µM of niclosamide analogues #1 and #2. The p values for the designated samples calculated by Student's t-test were <0.0001 (C) and 0.006 (D).