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Fig. 3 | Journal of Biomedical Science

Fig. 3

From: Degradation of neurodegenerative disease-associated TDP-43 aggregates and oligomers via a proteolysis-targeting chimera

Fig. 3

PROTAC 2 decreased insoluble C-TDP-43 aggregates via UPS. A Western blot of eGFP-C-TDP-43 harboring Neuro-2a cells with or without PROTAC 2 (5 μM) or/and MG132 (2 μM) treatment. The blots of RIPA-insoluble and RIPA-soluble cell lysates were demonstrated by SDS-PAGE and probed with C-TDP-43, GFP, phospho-TDP-43 (Ser409/410), and GAPDH antibodies. B–D Quantification of blots of C-TDP-43 (B), GFP (C), and phospho-TDP-43 (D) in panel A. E Representative images of mCherry-C-TDP-43 expressed Neuro-2a cells with or without PROTAC 2 (5 μM) or/and MG132 (2 μM). To morphologically monitor the C-TDP-43 puncta upon drug treatment, the cells with mCherry-C-TDP-43 puncta larger than 0.1 μm2 were considered as the aggregate-positive cells (featured with dash line). Scale bar = 10 μm. F Quantification of the percentage of aggregate-positive Neuro-2a cells among total cells in panel E. All the statistic results were quantified by ImageJ and shown as mean ± SD (n ≥ 3). Data were analyzed by one-way ANOVA with Tukey post-hoc test (*P < 0.05, **P < 0.01, ***P < 0.001)

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