There were errors in J. Cell Sci. (2008) 121, 2685-2695 (). The authors wish to correct errors in Figs 2B, 3B, 5A and 6A, and they have provided the original image data for these experiments.
In Fig. 2B, an image splice made to exclude a lane from the silver-stained gel was not marked in the figure. The corrected panel is shown below.
LC3 specifically interacts with p62 in an N-terminus-dependent manner. (B) Cell extracts (3.5 mg) prepared from GFP-LC3-expressing cells were precipitated using 3.5 μl anti-GFP antibodies. Samples were separated on 12% SDS-PAGE and stained using silver staining technique. M, Marker; E, eluate; T, total; F, flow through. The interacting protein (boxed band) was identified by mass spectrometry (MS).
LC3 specifically interacts with p62 in an N-terminus-dependent manner. (B) Cell extracts (3.5 mg) prepared from GFP-LC3-expressing cells were precipitated using 3.5 μl anti-GFP antibodies. Samples were separated on 12% SDS-PAGE and stained using silver staining technique. M, Marker; E, eluate; T, total; F, flow through. The interacting protein (boxed band) was identified by mass spectrometry (MS).
In Fig. 3B, image splicing to remove a lane from the western blots resulted in the appearance of some bands being altered, and the image splice was incorrectly applied to the anti-p62 western blot of cell extracts, resulting in mislabelling of lanes for that blot. The corrected panel, which uses an alternative film exposure of the original western blot, is shown below.
Two sites in LC3 are essential for p62 binding. HeLa cells were transfected with the gene of interest and after 24 hours were cultured for 2 hours in EBSS medium in the presence of 0.1 μM Baf A. Cells were then lysed using RIPA extraction buffer. GFP or GFP fusion constructs of LC3 and its mutants (A) or LC3 family proteins and their mutants (B) were immunoprecipitated (IP) from total cellular extract (500 μg) and subjected to SDS-PAGE. Copurified p62 was detected by immunoblotting with anti-p62 antibody (upper panel). WB, western blot. Immunoprecipitated GFP fusion proteins were detected with anti-GFP antibody (lower panel). Amount of total p62 or GFP fused proteins detected in 50 μg total protein extract (extracts) is indicated.
Two sites in LC3 are essential for p62 binding. HeLa cells were transfected with the gene of interest and after 24 hours were cultured for 2 hours in EBSS medium in the presence of 0.1 μM Baf A. Cells were then lysed using RIPA extraction buffer. GFP or GFP fusion constructs of LC3 and its mutants (A) or LC3 family proteins and their mutants (B) were immunoprecipitated (IP) from total cellular extract (500 μg) and subjected to SDS-PAGE. Copurified p62 was detected by immunoblotting with anti-p62 antibody (upper panel). WB, western blot. Immunoprecipitated GFP fusion proteins were detected with anti-GFP antibody (lower panel). Amount of total p62 or GFP fused proteins detected in 50 μg total protein extract (extracts) is indicated.
In Fig. 5A, an image splice to remove lanes was not marked, and the image splice was incorrectly applied to the anti-actin blot, resulting in mislabelling of lanes for that blot. The authors have confirmed that the western blot quantification in Fig. 5B is unaffected by these errors in figure assembly. The corrected panel for Fig. 5A is shown below.
LC3 is essential for starvation-induced degradation of p62. (A) HeLa cells were transfected with either transfection reagent (mock), nontargeting siRNA (control siRNA) or a pool of LC3 siRNAs (A, B and C isoforms) using DharmaFect reagent. After 72 hours, the cells were incubated in EBSS medium in the absence or presence of 0.1 μM Baf A, lysed with RIPA extraction buffer and analyzed by western blot with anti-p62, anti-actin and anti-LC3 antibodies. The asterisk indicates a nonspecific band. Arrows indicate LC3-I and LC3-II.
LC3 is essential for starvation-induced degradation of p62. (A) HeLa cells were transfected with either transfection reagent (mock), nontargeting siRNA (control siRNA) or a pool of LC3 siRNAs (A, B and C isoforms) using DharmaFect reagent. After 72 hours, the cells were incubated in EBSS medium in the absence or presence of 0.1 μM Baf A, lysed with RIPA extraction buffer and analyzed by western blot with anti-p62, anti-actin and anti-LC3 antibodies. The asterisk indicates a nonspecific band. Arrows indicate LC3-I and LC3-II.
In Fig. 6A, lanes from the anti-actin blot in Fig. 5A were incorrectly presented as the anti-actin control blot. Additionally, image splices were inconsistently applied in the anti-ubiquitin (UB) and anti-p62 blots, resulting in incorrect bands being shown for the ubiquitin control siRNA lane and both p62 lanes. The authors confirm that the western blot quantification in Fig. 6A is unaffected by these errors in figure assembly. The corrected panel is shown below.
Knockdown of LC3 leads to accumulation of detergent-insoluble p62 and polyubiquitylated proteins. (A) HeLa cells were transfected with either nontargeting siRNA (control siRNA), or with a pool of LC3 siRNAs (A, B and C isoforms) using DharmaFect reagent. After 72 hours, the cells were incubated in EBSS medium, lysed with RIPA extraction buffer and analyzed by western blot with anti-p62, anti-actin and anti-ubiquitin antibodies (left panel). Quantification of relative ubiquitin level (right panel) was performed as described in Materials and Methods. Mean±s.d. of three independent experiments is presented below.
Knockdown of LC3 leads to accumulation of detergent-insoluble p62 and polyubiquitylated proteins. (A) HeLa cells were transfected with either nontargeting siRNA (control siRNA), or with a pool of LC3 siRNAs (A, B and C isoforms) using DharmaFect reagent. After 72 hours, the cells were incubated in EBSS medium, lysed with RIPA extraction buffer and analyzed by western blot with anti-p62, anti-actin and anti-ubiquitin antibodies (left panel). Quantification of relative ubiquitin level (right panel) was performed as described in Materials and Methods. Mean±s.d. of three independent experiments is presented below.
The authors apologise to readers for these errors, which do not impact the results or conclusions of the article.
