Literature DB >> 26431316

Correction: Cysteine String Protein Limits Expression of the Large Conductance, Calcium-Activated K+ (BK) Channel.

Eva Ahrendt, Barry Kyle, Andrew P Braun, Janice E A Braun.   

Abstract

Entities:  

Year:  2015        PMID: 26431316      PMCID: PMC4591990          DOI: 10.1371/journal.pone.0140073

Source DB:  PubMed          Journal:  PLoS One        ISSN: 1932-6203            Impact factor:   3.240


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Fig 1C is incorrect as it shows the wrong actin blot. The authors have provided a corrected version of Fig 1 here.
Fig 1

CSPα alters BK channel expression.

A. Native CAD cells were transiently transfected with 1 μg cDNA encoding a neuronal BKα subunit, along with different amounts of myc-tagged CSPα (0.25 μg, 0.5 μg and 0.75 μg). Empty pCMV expression vector (0.75 μg) was co-transfected with 1 μg BKα subunit cDNA as a transfection control. 24 h post-transfection, the cells were lysed and the expression of BKα subunit and myc-tagged protein was analyzed by Western Blot. β-actin detection is shown to verify comparable sample loading. B. Histogram depicting quantification of BKα subunit levels in CAD cells co-transfected with increasing amounts of CSPα cDNA. Data are presented as mean ± SE of 5 similar experiments; *p<0.05 vs. pCMV vector control. C.Cells were transfected with 1 μg cDNA encoding BKα subunit along with either 0.25 μg or 0.75 μg of myc-tagged CSPα or 1 μg of pCMV. 24 h and 48 h post-transfection, BKα subunit expression was analyzed by Western Blot. D. Histograms depicting quantification of immunoreactive BKα subunit observed in the presence of co-transfected CSPα, as displayed in panel C. BKα subunit immunoreactivity detected at 48 h is expressed relative to the level of BKα subunit observed at 24 h; data are presented as mean ± SE of 4 similar experiments. Statistical significance was determined using one way ANOVA, *p<0.05; **p<0.01.

CSPα alters BK channel expression.

A. Native CAD cells were transiently transfected with 1 μg cDNA encoding a neuronal BKα subunit, along with different amounts of myc-tagged CSPα (0.25 μg, 0.5 μg and 0.75 μg). Empty pCMV expression vector (0.75 μg) was co-transfected with 1 μg BKα subunit cDNA as a transfection control. 24 h post-transfection, the cells were lysed and the expression of BKα subunit and myc-tagged protein was analyzed by Western Blot. β-actin detection is shown to verify comparable sample loading. B. Histogram depicting quantification of BKα subunit levels in CAD cells co-transfected with increasing amounts of CSPα cDNA. Data are presented as mean ± SE of 5 similar experiments; *p<0.05 vs. pCMV vector control. C.Cells were transfected with 1 μg cDNA encoding BKα subunit along with either 0.25 μg or 0.75 μg of myc-tagged CSPα or 1 μg of pCMV. 24 h and 48 h post-transfection, BKα subunit expression was analyzed by Western Blot. D. Histograms depicting quantification of immunoreactive BKα subunit observed in the presence of co-transfected CSPα, as displayed in panel C. BKα subunit immunoreactivity detected at 48 h is expressed relative to the level of BKα subunit observed at 24 h; data are presented as mean ± SE of 4 similar experiments. Statistical significance was determined using one way ANOVA, *p<0.05; **p<0.01. Additionally, there is a sentence missing from the caption for Fig 3. Please see the complete, correct Fig 3 caption here. The missing sentence is highlighted in bold.
Fig 3

The J domain of CSPα reduces BK channel expression.

A. Schematic of myc-tagged full length CSPα and CSPα deletion constructs. B. Western blot analysis of BK channel expression in CAD cells 24 h post-transfection with 1 μg cDNA encoding BKα subunit along with 0.75 μg myc-tagged full length CSPα cDNA or the indicated deletion constructs. As a transfection control, 0.75 μg empty pCMV was co-transfected with BKα subunit cDNA. 30 μg of cell lysate isolated under each experimental condition was separated by SDS-PAGE, probed with an anti-BKα subunit antibody and an anti-myc antibody. Data shown in 3B right panel is from the same blot; a lane between lanes 1 and 2 was removed. The histograms in panel C quantify changes in BK channel expression in the presence of wild-type CSPα and individual CSPα deletion mutants. Statistically significant differences from the pCMV control (set to 100%) were determined by one-way ANOVA; *p<0.05; **p<0.001.

The J domain of CSPα reduces BK channel expression.

A. Schematic of myc-tagged full length CSPα and CSPα deletion constructs. B. Western blot analysis of BK channel expression in CAD cells 24 h post-transfection with 1 μg cDNA encoding BKα subunit along with 0.75 μg myc-tagged full length CSPα cDNA or the indicated deletion constructs. As a transfection control, 0.75 μg empty pCMV was co-transfected with BKα subunit cDNA. 30 μg of cell lysate isolated under each experimental condition was separated by SDS-PAGE, probed with an anti-BKα subunit antibody and an anti-myc antibody. Data shown in 3B right panel is from the same blot; a lane between lanes 1 and 2 was removed. The histograms in panel C quantify changes in BK channel expression in the presence of wild-type CSPα and individual CSPα deletion mutants. Statistically significant differences from the pCMV control (set to 100%) were determined by one-way ANOVA; *p<0.05; **p<0.001.

Uncropped blots for Fig 1A, Fig 1C, and Fig 3B.

(PPTX) Click here for additional data file.
  1 in total

1.  Cysteine string protein limits expression of the large conductance, calcium-activated K⁺ (BK) channel.

Authors:  Eva Ahrendt; Barry Kyle; Andrew P Braun; Janice E A Braun
Journal:  PLoS One       Date:  2014-01-27       Impact factor: 3.240

  1 in total

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