Fig. 1

Characterization of TDP-43 expression patterns in aging and stroke. a Immunofluorescence of the brain cortex of wild-type 3-month-old mice using TDP-43 antibody at different time points after MCAO reveals mislocalization of TDP-43 protein into cytoplasm starting 72 h after MCAO. b Double immunofluorescence of the brain cortex sections of 3- and c 12-month-old mice in control conditions CTL (upper panels) and 72 h after MCAO using TDP-43 antibody (green) and NeuN antibody (red) show nuclear localization of TDP-43 in control conditions and cytoplasmic mislocalization of TDP-43 in neuronal cells in both age groups. d, e Western blot of nuclear lysates from 3- and 12-month-old mice using TDP-43 antibody in control and 72 h post MCAO does not reveal any significant changes in the levels of whole length TDP-43. P84 is used as loading control. f Western blot of cytoplasmic lysates from 3- and 12-month-old mice using TDP-43 antibody in control and 72 h after MCAO show expression of whole length TDP-43, fragmented TDP-35, and TDP-25. g Western blot of cytoplasmic lysates from 3- and 12-month-old mice using phospho-TDP-43 antibody in control and 72 h after MCAO show no expression of whole length P-TDP-43, fragmented P-TDP-35, or P-TDP-25. h Normalized densitometry values of immunoblots from control and 72 h after MCAO reveal significant increase in the levels of whole length TDP-43, pathological TDP-35, and TDP-25 fragments in aging. Actin is used as loading control. Quantified data in the figure was presented as mean ± SEM and statistical significance between the groups was achieved using ANOVA followed by Tukey’s multiple comparison test and depicted as ***p < 0.001. Scale bar represents 10 and 20 μm