Figure 6: LPS induces posttranslational modifications of TDP-43.

(a) RAW macrophages stably expressing HA-TDP-43 and GFP-TDP-43 were stimulated with LPS (80 ng ml⁻¹) for 6 h. Cell extracts were immunoprecipitated with anti-HA antibody and then subjected to immunoblot analysis with anti-TDP-43 antibody. TDP-43 expression was confirmed by immunoblot analysis using anti-HA, anti-GFP or anti-TDP-43 antibodies. (b) RAW macrophages were stimulated with LPS (80 ng ml⁻¹), followed by immunoprecipitation with rat anti-TDP-43 antibody and probing with mouse anti-TDP-43 antibody. Asterisks and arrowheads indicate the immunoglobulin heavy chains and cleaved TDP-43, respectively. (c) RAW macrophages were stimulated with LPS (80 ng ml⁻¹) and then treated with MG132 (10 μM). Cells were lysed with 2% NP-40, immunoprecipitated with anti-TDP-43 antibody and then analysed by immunoblotting with anti-ubiquitin antibody. Immunoprecipitated TDP-43 was confirmed by immunoblot analysis using anti-TDP-43 antibodies. Asterisk, ubiquitinated TDP-43. Data are representative of three independent experiments.