In the article by Zhou et al. “HSP72 Inhibits Smad3 Activation and Nuclear Translocation in Renal Epithelial-to-Mesenchymal Transition” (J Am Soc Nephrol. 2010; 21(4):598–609. DOI: 10.1681/ASN.2009050552), corrections are needed.
The authors wish to correct errors in two figures and provide clarifications in the figure legends. The corrections do not affect the scientific conclusions of the study.
During figure assembly, inadvertent errors occurred in the preparation of two Western blot panels.
In Figure 3B, the Smad3 panel inadvertently duplicated a band from Figure 3A. The panel has now been replaced with the correct original image from the same experiment.
In Figure 7H, the p-Smad3 panel inadvertently duplicated a band from Figure 6D. The panel has now been replaced with the correct original image from the same experiment.
The original images in Figures 3A and 6D are correct and remain unchanged.
The authors wish to clarify that certain figures were derived from the same original membranes, as they represent different target proteins assessed from identical experimental samples.
For Figures 2 and 5: Panels 2A and 5A were obtained from the same membranes and show epithelial-to-mesenchymal transition markers (Figure 2A) and Smad proteins (Figure 5A) from identical experimental samples. The HSP72 and β-actin bands are therefore the same in both panels.
For Figure 7: Panels 7A and 7D were obtained from the same membranes and represent different targets from identical experimental samples. The shared bands HSP72 and β-actin are therefore the same in both panels.
The authors confirm that all scientific conclusions of the article remain valid and are fully supported by the corrected data. The full corrected figures appear below.
Figure 3.
HSP72 suppresses activation of the TGF-β/Smads pathway. (A) Serum-deprived NRK-52E cells were treated with 10 ng/ml of TGF-β1 for the indicated time period. Cell lysates were probed with antibodies against p-Smad3, Smad3, p-Smad2, Smad2, Smad7 or β-actin. (B) NRK-52E cells were transfected with pcDNA3.1-HA-Wt-HSP72 (30 and 50 nM) or specific HSP72 siRNA were stimulated with 10 ng/ml of TGF-β1 for 30 minutes. (C–E) Smad protein content evaluated by Western blotting with densitometric analysis of the effect of HSP72 expression on p-Smad3, p-Smad2 and Smad7 content normalized with Smad2, Smad3, or β-actin content in TGF-β1-treated cells. Data are expressed as mean ± SEM; n = 3 per treatment; *P < 0.01 versus negative control; †P < 0.05 versus TGF-β1-treated cells without HSP72 overexpression or HSP72 siRNA.
Figure 7.
Endogenous HSP72 suppresses Smad signaling and EMT. (A) HK-2 cells transfected with pcDNA3.1-HA-Wt-HSP72 or specific HSP72 siRNA were exposed to 10 ng/ml of TGF-β1 for 48 hours. Western blotting was carried out as described in Figure 2A. (B and C) Graphic representation of densitometric quantification of the protein bands in panel A with E-cadherin or α-SMA normalized to the corresponding β-actin levels. Data are expressed as mean ± SEM; n = 3 per treatment; *P < 0.01 versus negative control; †P < 0.05 versus TGF-β1-treated cells alone. (D) Cells were treated and described as above. Expression of Smad2, Smad3, and Smad7 and phosphorylation of Smad2 and Smad3 were assayed by Western blot analyses. (E) Protein levels were quantified with scanning densitometry and normalized with Smad2, Smad3, or β-actin. Data are expressed as mean ± SEM of three independent experiments; *P < 0.01 versus negative control; †P < 0.05 versus TGF-β1-treated cells alone. (F–H) Coimmunoprecipitation of endogenous HSP72 and Smad3 or p-Smad3. Levels of HSP72, Smad3, and p-Smad3 in the IP products were analyzed by Western blotting.