Molecules and Cells

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Fig. 7.

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Fig. 7. Ana3 is secreted via exosomes and increases the level of exosomal Sona. In all western analyses, the fractions of cells are marked on the left, and the names and amounts (μg) of DNA constructs transfected to the S2 cells are given on the top of each panel. Antibodies used for immunoblots (IB) are indicated. (A) Ana3 is secreted via exosomes. The 252 kDa band indicates Ana3-GFP (arrow) and the 225 kDa band indicates endogenous Ana3 (arrowhead). Ana3 is detected in both P100 exosomal fraction and SNΔ fraction. (B) Ana3 stabilizes Sona. S2 cells were co-transfected with increasing amounts of ana3-GFP and a constant amount of sona-HA cDNA, and the levels of Sona in CX and cleared conditioned media (CM) were compared. The 75 kDa full-length Sona (arrows) was detected with Sona-Pro antibody or anti-HA antibody. The 37 kDa active Sona (arrowheads) was detected with anti-HA antibody. The 22 kDa cleaved pro-domain of Sona (asterisks) was detected by Sona-Pro antibody. (C) Ana3 significantly increased the level of Sona in P100 but not in SNΔ or CX fraction. The 75 kDa full-length Sona (arrow) and the 37 kDa active Sona (arrowheads) were detected with anti-HA antibody. The 37 kDa GAPDH and the 35 kDa Syntaxin 1A (Syx1A) were used as a cytoplasmic marker and an exosomal marker, respectively.
Mol. Cells 2021;44:13~25 https://doi.org/10.14348/molcells.2020.0244
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