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. 2016 Sep 27;139(12):3187–3201. doi: 10.1093/brain/aww237

Figure 1.

Figure 1

TDP-43 is secreted via exosomes in Neuro2a cells. (A) Electron microscopic image of secreted vesicles from Neuro2a cells. Scale bar = 100 nm. (B and C) Immunoelectron microscopic images with anti-CD63 (B) or Alix (C) antibodies. Scale bars = 100 nm. (D) Immunoblots of P100 and cell lysate (CL) of Neuro2a cells. (E) Immunoblots of pellets from different centrifugations of medium or cell lysates of Neuro2a cells transfected with wild-type (WT), A315T, G348C, or C-terminal fragment (25CTF) of hTDP-43. (F) Sucrose density gradient fractionation of P100 of Neuro2a cells transfected with V5-hTDP-43. Fifteen fractions of sucrose gradient, pellet (P), and cell lysate of Neuro2a cells. (G) Immunoelectron microscopic image of P100 of Neuro2a cells transfected with wild-type, A315T, G348C, or 25CTF hTDP-43 with V5 antibody. Scale bars = 100 nm. (H) Immunofluorescent images of Neuro2a cells transfected with V5-wild-type or -25CTF hTDP-43, Tsg101-Cherry (green; V5-TDP-43, red; Tsg101-Cherry, blue; DAPI). Dotted line indicates boundary of nucleus. Scale bars = 10 μm. Arrows indicate co-localization of V5-TDP-43 and Tsg101-Cherry. (I) Immunoblots of exosome fraction from primary astrocytes, primary microglial cells or primary neurons with brain lysate (BL) of C57BL6 mouse. Primary astrocytes or microglial cells were treated with DMSO or LPS (100 ng/ml) for 24 h.