[Journal] Nature Cell Biology. Volume 11. Number 4
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Additional resources for [Journal] Nature Cell Biology. Volume 11. Number 4
The percentage of colocalization between Atg14L and Atg16L or LC3-positive dot is shown. For example, (Atg14L×Atg16L)/Atg16L means the ratio of colocalization of Atg14L and Atg16L puncta to all of the Atg16L-positive puncta. d. of 25 cells. d. of 20 cells. d) Immunofluorescence with an anti-calnexin (ER) antibody. A549 cells transfected with adenovirus-based GFP–Atg14L. Scale bar, 10 μm. (e) Immuno-EM analysis of GFP–Atg14L. A549 cells transfected with adenovirus-vector based GFP or GFP–Atg14L were subjected to immuno-EM analysis using an anti-GFP antibody.
Oligonucleotide sequences for shRNA interference with Atg14L, UVRAG, or Rubicon expression are bp 383–405 of Atg14L (5´-GCAAGAUGAGGAUUGAACA‑3´), bp 513–536 of Rubicon (5´-GAUCGAUGCGUCCAUGUUU‑3´), bp 484–502 of UVRAG (5´-GCCAGACCGTCTTGATACA‑3´) and GL2 luciferase (negative control, 5´-CGTACGCGGAATACTTCGA‑3´) followed by a 9‑nucleotide non-complementary spacer (TTCAAGAGA) and the reverse complement of the initial 19-nucleotide sequence. These dsDNA oligonucleotides were cloned into the pENTR/U6 vector (Invitrogen) and transferred into the pAd/PL vector (Invitrogen) by the Clonase LR recombination reaction (Invitrogen).
J. Cell Biol. 171, 603–614 (2005). Mizushima, N. et al. Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells. J. Cell Biol. 152, 657–668 (2001). , Ueno, T. & Kominami, E. Lysosomal turnover, but not a cellular level, of endogenous LC3 is a marker for autophagy. Autophagy 1, 84–91 (2005). Mizushima, N. et al. Mouse Apg16L, a novel WD‑repeat protein, targets to the autophagic isolation membrane with the Apg12–Apg5 conjugate. J. Cell Sci. 116, 1679–1688 (2003). , Kameyama, K.