Figures
Due to a clerical error by the authors, Fig 1A of the original article includes overlapping images of the same cluster of cells in the "311" panel and in the "hypoxia" panel. In the revised version of Fig 1 provided here, all images in Fig 1A have now been replaced by replicate images obtained in parallel experiments. These replicate images show the same result as in the original Fig 1A, and demonstrate that a range of stressors (e.g., DFO, 311, hypoxia and tunicamycin) lead to the formation of eIF3a- containing structures consistent with stress-granules. The raw, uncropped immunofluorescence images for the revised figure are provided as a Supporting Information file.
(A) Cells were incubated using these conditions for 24 h/37°C and then stained with primary antibody against eIF3a (Alexa Fluor 555; “red”) and DAPI. Scale bar: 10 μm. (B) Co-localization of the stress granule markers eIF3a and eIF2α(Alexa Fluor 488; “green”) in structures consistent with stress granules after incubation with DFO under the conditions used in (A). The enlarged views of the boxed region in the merge panel are displayed to the right of this panel and show separate views for the red and green channels of the same field in which there is a cluster of eIF3a- and eIF2α-positive stress granules. The white arrows point to these structures. The scale bar in the left-most merge image represents 10 μm, while the scale bar in the enlarged merge image represents 2 μm. (C) Fractionation of MCF7 cells followed by western analysis demonstrated the presence of eIF3a and NDRG1 in both the cytoplasm and nucleus. GAPDH and HDAC1 were used as positive and loading controls for isolation of cytoplasmic (C) and nuclear (N) fractions, respectively. β-actin was used as general protein loading control. The blots are representative of 3 experiments and the densitometric analysis is expressed as mean ± SD. *p<0.05, **p<0.01, ***p<0.001 relative to the control of the same fraction.
Supporting Information
S1 File. Raw, uncropped immunofluorescence images for revised Fig 1A and 1B.
https://doi.org/10.1371/journal.pone.0149922.s001
(PPTX)
Reference
Citation: Lane DJR, Saletta F, Rahmanto YS, Kovacevic Z, Richardson DR (2016) Correction: N-myc Downstream Regulated 1 (NDRG1) Is Regulated by Eukaryotic Initiation Factor 3a (eIF3a) during Cellular Stress Caused by Iron Depletion. PLoS ONE 11(2): e0149922. https://doi.org/10.1371/journal.pone.0149922
Published: February 25, 2016
Copyright: © 2016 Lane et al. 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.