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Erratum: Characterization of calmodulin- free murine inducible nitric-oxide synthase (PLoS ONE (2015) 10: 3 (e0121782) DOI: 10.1371/journal.pone.0121782)
L. Nagpal,
Published in Public Library of Science
PMID: 33035256
Volume: 15
Issue: 10 October
The authors issue this Correction to address the following issues that were noted after this article [1] was published: • In Fig 1A, there appear to be vertical discontinuities between lanes, the lanes were mislabeled, and the image was stretched vertically from its original size. The authors commented that the apparent discontinuities resulted from a streaking effect observed between lanes on the original gel image. The original image supporting the published figure panel was not provided, but an updated Fig 1 is provided here, in which panel A is replaced with replicate data. The raw image supporting the updated version of Fig 1A is in S1 File, and additional replication data supporting this experiment are in S2 File. • Questions were raised about the lack of loading control data reported in [1] for the blots shown in Figs 2A, 2B, and 2D. Figs 2A-2D show results of western blot (A, B, D) and SDS-PAGE (C) experiments in which inputs were total bacterial extract (A) or affinity column- purified iNOS (B-D). The authors clarified that for western blot experiments, membranes were stained with Ponceau S after protein electro-transfer and prior to antibody probing both to confirm the efficacy of protein transfer as well as to be evaluated as loading controls. Ponceau S staining results and raw blot/gel image data for the reported experiments are provided in S3 File. As demonstrated in Fig 2A and S5 File, the level of iNOSfl was observed to be substantially lower when the protein was expressed in the absence of CaM than in its presence. Higher levels of degraded iNOSfl proteins were observed in the iNOSfl preparations expressed without CaM. Equal amounts of lysate were loaded in lanes 1 & 2 for this experiment. For Figs 2B-2D, different amounts of purified iNOSfl proteins were loaded in the plus versus minus CaM lanes so as to obtain approximately equivalent iNOSfl levels (130 KD bands) across lanes within each gel or blot. Loading volumes needed to achieve equivalent iNOSfl levels and were determined through standardization experiments for which data were not reported in the article. The iNOS blots in Fig 2B and 2D served as the controls for the CaM blot in Fig 2B and the heme gel in Fig 2C, respectively. For each of these experiments, duplicate gels were prepared for the control and experimental assays by loading the same purified iNOSfl proteins on parallel gels. • In Fig 3, the iNOSfl western blot in panel B1 errantly reports a horizontally flipped version of the western blot in panel B2. The authors note that this was due to a figure assembly error. This is corrected in the updated version of Fig 3 provided here. The original blots underlying these panels are provided in S4 and S5 Files. (Figure presented) • The article’s Data Availability statement says, “All relevant data are within the paper.” The underlying data were not included with [1] but are provided herein S1–S9 Files. All original data are available except those supporting Fig 2D. A member of PLOS ONE’s Editorial Board reviewed the updated figures, supporting data files, and information provided above regarding western blot controls and advised that they address the previous concerns and errors in the original manuscript. The Academic Editor commented that the corrections do not change the article’s conclusions. The authors apologize for the errors in the published article. Supporting information S1 File. Original image supporting the replacement results shown in the revised version of Fig 1A. (PPT) S2 File. Replicate data supporting Fig 1A. (TIF) S3 File. Underlying data supporting Fig 2. The blot image provided for Fig 2D is a digital image of the original blot reported in the published figure; levels were adjusted in the image file so that band intensities would align approximately with bands observed by the Ponceau S staining. The original film from the Fig 2D blot experiment is no longer available. (ZIP) S4 File. Original image data supporting Fig 3B1. (TIF) S5 File. Original image data supporting Fig 3B2. (JPG) S6 File. Underlying data supporting Fig 3A1, 3A2, 3C1 and 3C2. (ZIP) S7 File. Raw data supporting Fig 6. (PDF) S8 File. Raw data supporting Table 1. (ZIP) S9 File. Raw data supporting Table 2. (ZIP). © 2020 Nagpal, Panda. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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JournalPLoS ONE
PublisherPublic Library of Science