Pbs dapi

The sections were washed in PBS (3 times, 5 minutes each), permeabilized and blocked in a PBS/0.3% Tween 20/1.5% BSA for one hour at room temperature, followed by an overnight incubation with the primary antibody diluted in a PBS/0.2% TritonX100 solution at room temperature. Next, sections were washed in PBS (3 times, 10 minutes each) and incubated with the secondary antibody (diluted in PBS) for one hour, washed twice with PBS (10 minutes each), and then incubated for 10 minutes with PBS/DAPI (Thermo Fisher Scientific, 100ng/ml) solution. To quench autofluorescence, sections were incubated with 0.1% Sudan Black/70% EtOH for 1 minute, and then washed twice briefly with PBS. Sections were mounted on Fisherbrand Superfrost Plus Microscope Slides (Thermo Fisher Scientific) with ProLong Gold antifade reagent with DAPI (Thermo Fisher Scientific).

The sections were washed in PBS (3 times, 5 minutes each), permeabilized and blocked in a PBS/0.5% Tween 20/1.5% BSA for one hour at room temperature, followed by an overnight incubation with the primary antibody anti-TDP-43 (12892–1-AP, Proteintech; 1:500) diluted in a PBS/0.3% TritonX100 solution at room temperature. Next, sections were washed in PBS (3 times, 10 minutes each) and incubated with the secondary antibody (diluted in PBS) for one hour, washed twice with PBS (10 minutes each), and then incubated for 10 minutes with PBS/DAPI (Thermo Fisher Scientific, 100ng/ml) solution. Sections were mounted on Fisherbrand Superfrost Plus Microscope Slides (Thermo Fisher Scientific) with ProLong Gold antifade reagent with DAPI (Thermo Fisher Scientific).