Fig. 2

Cytoplasmic TDP-43 shows ubiquitin positivity and microglia/macrophages exhibit TDP-43 mislocalization. a Double immunofluorescence of the brain cortex sections of 3- and 12-month-old mice 72 h after MCAO using TDP-43 antibody (green) and ubiquitin (red) reveal the formation of cytoplasmic TDP-43-ubiquinated aggregates 72 h after MCAO in 3- and 12-month-old mice. b Co-immunoprecipitation of ubiquitin using rabbit polyclonal TDP-43 from the cytoplasmic lysates of 3- and 12-month-old control and MCAO operated mice reveal the ubiquitination of TDP-43 after 72 h post MCAO. Note that TDP-43 ubiquitination is increased in 12-month-old transgenic mice after MCAO. Western blot of TDP-43 using rabbit polyclonal TDP-43 is shown as 15% input to confirm the presence of TDP-43 protein in the cytoplasmic lysate we used for Co-immunoprecipitation and actin is shown as loading control. c Double immunofluorescence of the brain cortex sections of 3- and 12-month-old mice 72 h after MCAO using TDP-43 antibody (red) and CD11b antibody (green) show mislocalization of TDP-43 in microglial cells in both age groups. Scale bar represents 10 μm