Fig. 2

CLU inhibits TDP-43 aggregation and reduces the number of TDP-43-GFP inclusions in neuronal cells during ER stress. a TDP-43^286-331 (peptide) was incubated at 37 °C with shaking, in the presence of either CLU or the non-chaperone control protein bovine superoxide dismutase (SOD1), at the molar ratios indicated in the key. Peptide aggregation was monitored by ThioT fluorescence. Data are means +/−SE (n = 4). b Upper panel: Image of Western blot detecting soluble TDP-43-tGFP after incubation at 4 °C, or 37 °C (which increases aggregation), with or without CLU or BSA (a non-chaperone control protein). Black arrow indicates position of monomeric TDP-43-tGFP, upper bands represent soluble oligomers. Loading control (colored dye from Promega transcription/translation kit) shown at bottom. Lower panel: Densitometric quantification of blot results (means + SD, n = 3). c Transfected N2a cells were treated as indicated for 10 h (key below plot), then lysed and TDP-43^M337V-tGFP inclusions enumerated by flow cytometry; data plotted as % change relative to untreated cells (means + SEM, n = 4; untreated cells are defined as the 100% value). See also Additional file 1: Figure S5. For all panels, results shown are representative of two or more independent experiments. In (b) and (c), horizontal bars indicate pairwise comparisons, * p < 0.05, ** p < 0.01, *** p < 0.001 (1-way ANOVA with Newman-Keuls post-test (b), or 2-way ANOVA with a Bonferroni post-test (c))