Quantitative Western Blot Analysis

A 15 μg aliquot of protein per sample was resolved on a denaturing 10% polyacrylamide gel, and proteins were transferred to polyvinylidene difluoride (PVDF) membranes using a semidry blotter by established standard protocols (Figueroa-Romero et al., 2009 (link); Lunn et al., 2009 (link)). Blots were probed overnight at 4°C with the following primary antibodies: rabbit anti-TDP43 (1:1,000; cat#A260, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-histone 2B (H2B) member S (1:500; cat# NB100-56347, Novus Biologicals, Littleton, CO, USA), rabbit anti-GFP (1:1,000; cat#2555, Cell Signaling Technology), and rat anti-α-tubulin (1:5,000; cat#ab6160, Abcam). The next day, blots were rinsed and probed for 50 min at room temperature with appropriate HRP-conjugated secondary antibodies (1:1,000; Santa Cruz Biotechnology or New England Biolabs), then rinsed and visualized by enhanced chemiluminescence with Prime Western Blotting Detection Reagent (Amersham, GE Healthcare, Chicago, IL, USA).