Skip to main content
. 2019 Nov 18;8:e48255. doi: 10.7554/eLife.48255

Figure 7. Human CLU_45 kDa is translated from an AUG start site in Exon 3.

Figure 7.

(A) Empty vector (pCMV6), V5-tagged pCMV6-human CLU Exon 2–9, or Exon 3–9 constructs were transfected into human SH-SY5Y cells. Cell lysates were harvested and probed for CLU immunoreactivity using anti-V5 (left panel) or anti-Clusterin (right panel). (B: left panel) pCMV6-human CLU Exon 2–9 or Exon 3–9 constructs were transfected into human SH-SY5Y cells. The resulting cell lysates were treated with PNGase F or Endo H and probed for CLU immunoreactivity using anti-V5. (B: right panel) Cell lysates harvested from pCMV6-human CLU Exon 3–9-transfected SH-SY5Y cells were treated with O-glycosidase, neuraminidase, or a combination of the two and probed for expression of CLU_45 kDa protein using anti-V5. (C) pCMV6-human CLU Exon 3–9-transfected SH-SY5Y cells were subjected to cellular fractionation using the Qiagen Qproteome Mitochondrial Isolation Kit and analyzed for CLU immunoreactivity using anti-V5. Fraction isolation was confirmed using calnexin (ER), Gapdh (cytosol), lamin A/C and lamin B1 (nucleus), and VDAC and Tom20 (mitochondria).