Higher levels of TDP-43 protein in LPS-activated astrocytes and microglia.

<p>Cultured astrocytes and microglia from transgenic (hTDP-43^A315T) and non-transgenic litters were treated for 1 day with LPS at 1 μg/ml or 0.1 μg/ml. Total protein was extracted from LPS -treated and untreated cultures and analysed by immunoblotting. TDP-43 was detected using the polyclonal antibody (Proteintech # 10782-AP) which reacts with both human (transgenic) and mouse (endogenous) proteins. TDP-43 bands were normalized against actin to take into account the difference in protein loading. The exposure times are actually different between transgenic and non-transgenic blots. Due to higher quantities of TDP-43 (human plus mouse) in the transgenic cell cultures the exposure time was only 30 seconds whereas for the non-transgenic cultures it was 4 minutes. Fold change is the ratio of band intensity in LPS-treated cultures over the band intensity in the respective untreated control. (A) Representative western blot of TDP-43 from LPS treated or untreated astrocyte culture. (B) Quantitative analysis of western blot showed that levels of total TDP-43 was higher in LPS treated transgenic astrocyte culture than non-transgenic culture. (C) Representative western blot of TDP-43 from LPS treated or untreated microglia culture. (D) Quantitative analysis of western blot showed that levels of total TDP-43 were also higher in LPS-treated microglia from transgenic TDP-43^A315T mice than from C57BL6 mice.