Cell Biology Correction for “β-Arrestin2 is a critical component of the GPCR–eNOS signalosome,” by Songling Liu, Louis M. Luttrell, Richard T. Premont, and Don C. Rockey, which was first published May 13, 2020; 10.1073/pnas.1922608117 (Proc. Natl. Acad. Sci. U.S.A. 117, 11483–11492).
The authors note that Fig. 5 appeared incorrectly.
Fig. 5.
ET-1 activation of β-Arr2 and eNOS. (A) SECs from normal (Left two panels) and BDL injured (Right two panels) rat livers were cultured for 24 h and exposed to ET-1 (20 nM) for 30 min. Cells were fixed and labeled with antibody to β-Arr2 (Upper red) and nuclei were labeled with DAPI (Lower, blue). Representative images (of more than 10 others) of single and merged channels are shown. (Scale bars, 10 μm.) (B) SECs isolated from normal rat livers (Left two panels) and BDL injured (Right two panels) rat livers were cultured for 24 h and exposed to ET-1 as in A. β-Arr2 and eNOS were immmunolabeled with anti–β-Arr2 and anti-eNOS antibodies. β-Arr2 (Alexa Fluor 647, green) and eNOS (Alexa Fluor 568, magenta) were visualized in horizontal cross-section (X-Y), in cross-section (X-Z) with an SR-200 inverted microscope (Vutara, Inc.) (60×/1.2 NA Olympus water immersion objective). Samples were imaged based on the SML biplane FPALM technology as described. Representative X-Y section (Upper) and X-Z (Bottom) section images from control (Left) and exposed to ET-1 (Right) are shown. (Scale bar, 1 μm.) (C) β-Arr2 and eNOS colocalization in normal and injured SECs as in B was quantified as in Materials and Methods; the changes in normal and injured SECs are presented graphically (1 for complete and 0 for no colocalization [n = 3/group]). (D) SECs isolated from β-Arr2 WT and KO mice were exposed to ET-1 (20 nM) for 30 min. Cell lysates were subjected to immunoblotting with the indicated antibodies. Specific bands corresponding to P-eNOS were quantified and are presented as the ratio of P-eNOS to eNOS, shown on the Right (n = 3/group). (E) eNOS enzymatic activity was measured in cell lysates treated as in D and NOS activity was normalized to that of control cells from β-Arr2 WT mice without ET-1 exposure and is presented graphically (the activity in SECs from β-Arr2 WT mice without ET-1 exposure was set at 100; n = 3/group). Statistical significance for C–E was evaluated by ordinary oneway ANOVA. Data are mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.005 for differences between indicated groups; n.s., no significant difference.
In Fig. 5B, the same image was mistakenly shown in the bottom Normal “Control” panel and in the bottom Injured “ET-1” panel. The corrected figure and its legend appear below. The online version has been corrected.
In Fig. 5D, the authors mistakenly selected an incorrect β-Actin blot for the experiment. In addition, there was a duplication in the bands of the β-Actin blot presented in the figure. This mistake has no bearing on the interpretation of the data for this experiment and has no effect on the conclusions presented in the manuscript.
In addition, the authors note that Supporting Fig. 4 in the SI Appendix appeared incorrectly. The published Fig. S4D has a mistake in the β-Actin panel. The original β-Actin blot included 8 lanes representing actin protein found in lanes in experiments shown in Fig. 1A and in Fig. S4D. The first 4 lanes were correctly taken from the blot and assigned to the experiment in Fig. 1A. However, lanes 4, 5, and 6 were accidently placed into Fig. S4D. The correct actin bands for this figure should have been lanes 6, 7 and 8. This mistake has no bearing on the interpretation of the data for this experiment and has no effect on the conclusions presented in the manuscript.
The SI Appendix has been corrected online.
Data Availability.
All relevant associated protocols, references to specific materials, and relevant data are contained within the manuscript and its SI Appendix files. To allow others to replicate data and/or build on the published work, the authors will provide readers access to our associated protocols, certain materials, and data from the study upon request.