Correction to "Increased expression of desmin and vimentin reduces bladder smooth muscle contractility via JNK2".

Javed E, Thangavel C, Frara N, et al. Increased expression of desmin and vimentin reduces bladder smooth muscle contractility via JNK2. The FASEB Journal. 2020;34:2126-2146. https://doi.org/10.1096/fj.201901301R The authors report that in Figure 2C and Figure 2D, identical bands are shown for GFP and GAPDH as these protein bands were obtained from the same experiment. The authors revised Figure 2C and Figure 2D using GFP and GAPDH bands from different experiments to avoid confusion. In addition, in Figure 2C, the bands labeled as GFP and GAPDH were misidentified while making the figure. This mistake has been corrected. The same GFP panel was used in Figures 6 and 7 because it was the control for desmin and vimentin overexpression. The figure legends have been corrected to make this clarification. In Figure 8, murine bladder smooth muscle (BSM) strips overexpressing GFP, or desmin, or vimentin were immunoblotted with antibodies against pJNK and GAPDH. BSM strips expressing GFP served as the control. The membrane was developed with short (Panel A) and long (Panel B) exposure times. The correct bands for pJNK and GAPDH were taken from the membrane with long-exposure time from GFP and vimentin (Figure 8E). However, the authors mistakenly included the bands for pJNK and GAPDH from the membrane with short exposure in Figure 8C from GFP and vimentin, instead of the intended bands from GFP and desmin. Figure 8C should have the pJNK and GAPDH bands from GFP and desmin. This error occurred due to the strong similarity between the pJNK bands in GFP/desmin and GFP/vimentin, as well as the GAPDH bands in GFP/desmin and GFP/vimentin. In the corrected Figure 8C, the bands for pJNK and GAPDH are included from GFP and desmin blots. The pJNK and GAPDH bands could come from different exposure times because they are from the same experiment. These errors do not change the results or conclusions of the article. The authors apologize for these errors. The corrected Figure 2 is as follows: FIGURE 2. Adenovirus-mediated overexpression of vimentin in murine BSM strips. (A) Schematic depiction of vimentin-GFP bicistronic adenoviral vector construct. (B) Schematic depiction of GFP adenoviral vector construct. (C, D) Murine BSM strips devoid of urothelium, and submucosa were transduced with an adenovirus encoding GFP and vimentin for 48 h and the expression levels of vimentin, desmin, and GFP proteins were determined by immunoblot analysis. GAPDH was used as a loading control. (E, F) Quantification of immunoblot data. (G) Sections prepared from murine BSM strips overexpressing vimentin and GFP proteins were stained with anti-vimentin, anti-SM22, and Alexa Fluor 488-labeled GFP antibody, followed by Cy3 and Cy5 conjugated secondary antibodies. Representative confocal images are shown. Scale bars = 10 μm. (H) Quantification of confocal images data. Data are expressed as means ± SD (E, F, and H), n = 5 mice in each group (E, F, and H). **p < .01 versus GFP-expressing murine BSM strips. FIGURE 6. Effect of desmin overexpression on smooth muscle marker proteins' expression in murine BSM. (A) Murine BSM strips devoid of urothelium, and submucosa were transduced with an adenovirus encoding GFP and desmin for 48 h and the expression levels of SMA and SM22 were determined by immunoblot analysis. GAPDH was used as a loading control. (B) Quantification of immunoblot data. (C, D) Sections prepared from murine BSM strips overexpressing desmin and GFP proteins were stained with anti-SMA, or anti-SM22, or anti-SMHC or Alexa Fluor 488-labeled GFP antibody, followed by Cy3 and Cy5 conjugated secondary antibodies. Representative confocal images are shown in (C) and (D); Scale bars = 50 μm. € Quantification of confocal images data. Data are expressed as means ± SD, n = 5 mice in each group. NS indicates nonsignificant. Note that the same GFP panel was used in Figures 6 and 7 because it was the control for both desmin and vimentin overexpression. FIGURE 7. Effect of vimentin overexpression on smooth muscle marker proteins' expression in murine BSM. (A) Murine BSM strips devoid of urothelium, and submucosa were transduced with an adenovirus encoding GFP and vimentin for 48 h and the expression levels of SMA, and SM22 were determined by immunoblot analysis. GAPDH was used as a loading control. (B) Quantification of immunoblot data. (C, D) Sections prepared from murine BSM strips overexpressing vimentin and GFP proteins were stained with anti-SMA, or anti- SM22, or anti-SMHC or Alexa Fluor 488-labeled GFP antibody, followed by Cy3 and Cy5 conjugated secondary antibodies. Representative confocal images are shown in C and D, Scale bars = 50 μm. (E) Quantification of confocal images data. Data are expressed as means ± SD, n = 5 mice in each group. NS indicates nonsignificant. Note that the same GFP panel was used in Figures 6 and 7 because it was the control for both desmin and vimentin overexpression. The corrected Figure 8 is as follows: FIGURE 8. Desmin and vimentin interact with JNK2 and enhance the phospho JNK level following the IF protein overexpression in murine BSM. (A, B) Murine BSM strips devoid of urothelium, and submucosa were transduced with an adenovirus encoding GFP, desmin and vimentin for 48 h and the total proteins from GFP, desmin and vimentin overexpressing murine BSM strips were immunoprecipitated with JNK2 antibody. The resulting immunoprecipitate was separated on a SDS-PAGE and subsequently probed with either anti-desmin and anti-JNK2 antibodies (A) or anti-vimentin and anti-JNK2 antibodies (B). The input served as a loading control in both (A) and (B). (C-F) Increased levels of phospho JNK following desmin and vimentin overexpression. Total proteins extracted from GFP, desmin and vimentin overexpressing murine BSM strips were subjected to immunoblot analysis using phosphorylated JNK and total JNK2 antibodies. GAPDH was used as a loading control. (D, F) Quantification of immunoblots (C, E). Data are expressed as means ± SD, n = 5 mice in each group. **p < .01 versus GFP expressing murine BSM strips.