Fig. 1

ALS-CSF induced NF-κB activation and TDP43 mislocalization. (a) Increased NF-κB activity in NSC-34 cells exposed to ALS-CSF (n = 3 in triplicates). (b, c). TDP43 analysis in-vitro. Representative image panels (b) and graph (c) showing cellular location of TDP43 in the NSC-34 cells across the study groups. Note the increased cytoplasmic TDP43 (red asterisks) in the ALS panel (n = 3, each in triplicates). Scale bar = 20 μm. (d- k) TDP43 immunodetection in-vivo in hTDP43 transgenic mice infused i.c.v. with PBS, CSF from non-ALS (NALS) or ALS. Representative image panels and graph for immunodetection of human TDP43 (d, f) and of pan-TDP43 recognizing both human and mouse species (e, g). Note neuronal (d, arrowhead) and glial (d, asterisk) TDP43 cytoplasmic mislocalization in the ALS-CSF panel. (h) represents qualitative and quantitative observations for the immunoblots of TDP43 obtained from the insoluble fraction of lysates using a pan-TDP43. Since the TDP43^WT construct in the transgenic mice was tagged with HA, the bands for human TDP43 appeared ~ 2 kDa on top of the endogenous mouse TDP43 (~ 43 kDa) on the immunoblot (i) represents qualitative and quantitative observations, of the immunoblots stained for the phosphorylated and non-phosphorylated forms of human (~ 45 kDa) and mouse (~ 43 kDa) TDP43. (n = 3, in triplicates) Scale bar = 10 μm. Data are mean ± SEM. (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 and **** p ≤ 0.001) and fold changes are calculated compared to NALS