Structured Review

Abcam anti kchip2
A , diagram for Flag-Kv4.2-GFP oriented in the plasma membrane. Flag and GFP were tagged in the extracellular S1-S2 loop and the C-terminal, respectively. B , Western blotting analysis of HEK293 cells expressing Kv4.2 with or without the expression of <t>KChIP2.</t> The plasmids transfected in HEK293 cells were indicated above each lane. The detecting antibodies were indicated at the right side. Lane 1 showed the lysate from cells expressing the corresponding vector as control. Lane 2 and 3 represented cells expressing Kv4.2 alone or with KChIP2, respectively. C , an example of I Kv4.2 recorded in HEK293 cells expressing Flag-Kv4.2-GFP. No significant difference of I Kv4.2 was found between Flag-Kv4.2-GFP-expressing cells and cells expressing untagged Kv4.2 (not shown). D , confocal images of HEK293 cells transfected with Flag-Kv4.2-GFP, showing subcellular localization of Kv4.2, with (P (+)) or without (P (−)) membrane permeabilization. The surface population of Kv4.2 was visualized by anti-Flag antibody ( red ) and the whole Kv4.2 was indicated by GFP ( green ). The nuclei were stained by diamidino-phenyl-indole (DAPI) ( blue ). Note that anti-Flag antibody detected only the membranous Kv4.2 when without permeabilization, while it detected the overall Kv4.2 after permeabilization. The merged images showed perfect co-localization of Flag-Kv4.2-GFP detected by anti-Flag antibody and GFP, respectively. Scale bars = 50 µm.
Anti Kchip2, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti kchip2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti kchip2 - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "Multi-Walled Carbon Nanotubes Impair Kv4.2/4.3 Channel Activities, Delay Membrane Repolarization and Induce Bradyarrhythmias in the Rat"

Article Title: Multi-Walled Carbon Nanotubes Impair Kv4.2/4.3 Channel Activities, Delay Membrane Repolarization and Induce Bradyarrhythmias in the Rat

Journal: PLoS ONE

doi: 10.1371/journal.pone.0101545

A , diagram for Flag-Kv4.2-GFP oriented in the plasma membrane. Flag and GFP were tagged in the extracellular S1-S2 loop and the C-terminal, respectively. B , Western blotting analysis of HEK293 cells expressing Kv4.2 with or without the expression of KChIP2. The plasmids transfected in HEK293 cells were indicated above each lane. The detecting antibodies were indicated at the right side. Lane 1 showed the lysate from cells expressing the corresponding vector as control. Lane 2 and 3 represented cells expressing Kv4.2 alone or with KChIP2, respectively. C , an example of I Kv4.2 recorded in HEK293 cells expressing Flag-Kv4.2-GFP. No significant difference of I Kv4.2 was found between Flag-Kv4.2-GFP-expressing cells and cells expressing untagged Kv4.2 (not shown). D , confocal images of HEK293 cells transfected with Flag-Kv4.2-GFP, showing subcellular localization of Kv4.2, with (P (+)) or without (P (−)) membrane permeabilization. The surface population of Kv4.2 was visualized by anti-Flag antibody ( red ) and the whole Kv4.2 was indicated by GFP ( green ). The nuclei were stained by diamidino-phenyl-indole (DAPI) ( blue ). Note that anti-Flag antibody detected only the membranous Kv4.2 when without permeabilization, while it detected the overall Kv4.2 after permeabilization. The merged images showed perfect co-localization of Flag-Kv4.2-GFP detected by anti-Flag antibody and GFP, respectively. Scale bars = 50 µm.
Figure Legend Snippet: A , diagram for Flag-Kv4.2-GFP oriented in the plasma membrane. Flag and GFP were tagged in the extracellular S1-S2 loop and the C-terminal, respectively. B , Western blotting analysis of HEK293 cells expressing Kv4.2 with or without the expression of KChIP2. The plasmids transfected in HEK293 cells were indicated above each lane. The detecting antibodies were indicated at the right side. Lane 1 showed the lysate from cells expressing the corresponding vector as control. Lane 2 and 3 represented cells expressing Kv4.2 alone or with KChIP2, respectively. C , an example of I Kv4.2 recorded in HEK293 cells expressing Flag-Kv4.2-GFP. No significant difference of I Kv4.2 was found between Flag-Kv4.2-GFP-expressing cells and cells expressing untagged Kv4.2 (not shown). D , confocal images of HEK293 cells transfected with Flag-Kv4.2-GFP, showing subcellular localization of Kv4.2, with (P (+)) or without (P (−)) membrane permeabilization. The surface population of Kv4.2 was visualized by anti-Flag antibody ( red ) and the whole Kv4.2 was indicated by GFP ( green ). The nuclei were stained by diamidino-phenyl-indole (DAPI) ( blue ). Note that anti-Flag antibody detected only the membranous Kv4.2 when without permeabilization, while it detected the overall Kv4.2 after permeabilization. The merged images showed perfect co-localization of Flag-Kv4.2-GFP detected by anti-Flag antibody and GFP, respectively. Scale bars = 50 µm.

Techniques Used: Western Blot, Expressing, Transfection, Plasmid Preparation, Staining

A and B , examples of activation ( A ) and inactivation ( B ) currents from transfected HEK293 cells. C and D , Boltzmann equation-fitted activation and inactivation curves, respectively. E , I Kv4.2 obtained by depolarizing the cell from −90 mV to +50 mV, showing accelerated decay upon MWCNTs treatment in HEK293 cells co-expressing Kv4.2 with KChIP2. F , statistical data of decay time constants (τ decay ) in various conditions. Abbreviations: C, carboxylated MWCNTs; P, pristine MWCNTs; A, aminated MWCNTs. ** P <0.01 vs. Kv4.2. ## P <0.01 vs. KChIP2.
Figure Legend Snippet: A and B , examples of activation ( A ) and inactivation ( B ) currents from transfected HEK293 cells. C and D , Boltzmann equation-fitted activation and inactivation curves, respectively. E , I Kv4.2 obtained by depolarizing the cell from −90 mV to +50 mV, showing accelerated decay upon MWCNTs treatment in HEK293 cells co-expressing Kv4.2 with KChIP2. F , statistical data of decay time constants (τ decay ) in various conditions. Abbreviations: C, carboxylated MWCNTs; P, pristine MWCNTs; A, aminated MWCNTs. ** P <0.01 vs. Kv4.2. ## P <0.01 vs. KChIP2.

Techniques Used: Activation Assay, Transfection, Expressing

A , I-V curves of I Kv4.2 at different conditions. B , the voltage-dependent activation and inactivation curves fitted with Boltzmann equation. C and D , effects of MWCNTs on the decay kinetics of HEK293 cells expressing Kv4.2 with or without KChIP2. ** P <0.01 vs. control. E and F , the recovery curve and the statistical recovery time constant (τ recovery ), respectively. ** P <0.01 vs. control.
Figure Legend Snippet: A , I-V curves of I Kv4.2 at different conditions. B , the voltage-dependent activation and inactivation curves fitted with Boltzmann equation. C and D , effects of MWCNTs on the decay kinetics of HEK293 cells expressing Kv4.2 with or without KChIP2. ** P <0.01 vs. control. E and F , the recovery curve and the statistical recovery time constant (τ recovery ), respectively. ** P <0.01 vs. control.

Techniques Used: Activation Assay, Expressing

A , Western blotting analysis showing changes of Kv4.2 and KChIP2 in two cell lines. B , statistical histograms from showing the fold changes after 6 h-treatment with MWCNTs. C , biotinylation results showing the surface population of Kv4.2, with the expression of endogenous Na + /K + -ATPase (NKP) as loading control. D , quantification histograms indicating the fold changes of the surface population of Kv4.2. * P <0.05 vs . control.
Figure Legend Snippet: A , Western blotting analysis showing changes of Kv4.2 and KChIP2 in two cell lines. B , statistical histograms from showing the fold changes after 6 h-treatment with MWCNTs. C , biotinylation results showing the surface population of Kv4.2, with the expression of endogenous Na + /K + -ATPase (NKP) as loading control. D , quantification histograms indicating the fold changes of the surface population of Kv4.2. * P <0.05 vs . control.

Techniques Used: Western Blot, Expressing

A , an original recording of flow cytometry showing the fluorescence of Flag which represented the population of Kv4.2 in cell membrane. A leftward shift of the trace meant a reduction of fluorescence intensity in membrane and vise versa. Untransfected HEK293 cells served as negative control. B , ratio of mean fluorescence intensity of Flag to GFP in HEK293 cells expressing Kv4.2 with or without KChIP2 upon MWCNTs treatment from . C , aliquots of cell lysate from HEK293 cells expressing Kv4.2 and KChIP2 were mixed with either anti-Kv4.2 antibody or anti-KChIP2 antibody and precipitated with protein A-sepharose. Immune complexes were resolved by SDS-PAGE. The immunoprecipitating antibodies were indicated above each lane, and detecting antibodies were shown at the left side. D , effects of MWCNTs treatment on the ratio of KChIP2 to Kv4.2 with anti-Kv4.2 antibody as bait for co-IP in HEK293 cells and cell lysate. MWCNTs were applied to the culture medium and cell lysate for 6 h, respectively. E , statistical results of the effects of MWCNTs on the ratio of KChIP2 to Kv4.2 in HEK293 cells expressing Kv4.2 and KChIP2. * P <0.05, ** P <0.01 vs. control. F , confocal images showing the membranous and intracellular distributions of Kv4.2. GFP ( green ) and anti-Flag (red ) represented the total and membranous Kv4.2, respectively. Note that MWCNT-treated cells showed reduced membranous Kv4.2 compared with control cells. Scale bars = 50 µm.
Figure Legend Snippet: A , an original recording of flow cytometry showing the fluorescence of Flag which represented the population of Kv4.2 in cell membrane. A leftward shift of the trace meant a reduction of fluorescence intensity in membrane and vise versa. Untransfected HEK293 cells served as negative control. B , ratio of mean fluorescence intensity of Flag to GFP in HEK293 cells expressing Kv4.2 with or without KChIP2 upon MWCNTs treatment from . C , aliquots of cell lysate from HEK293 cells expressing Kv4.2 and KChIP2 were mixed with either anti-Kv4.2 antibody or anti-KChIP2 antibody and precipitated with protein A-sepharose. Immune complexes were resolved by SDS-PAGE. The immunoprecipitating antibodies were indicated above each lane, and detecting antibodies were shown at the left side. D , effects of MWCNTs treatment on the ratio of KChIP2 to Kv4.2 with anti-Kv4.2 antibody as bait for co-IP in HEK293 cells and cell lysate. MWCNTs were applied to the culture medium and cell lysate for 6 h, respectively. E , statistical results of the effects of MWCNTs on the ratio of KChIP2 to Kv4.2 in HEK293 cells expressing Kv4.2 and KChIP2. * P <0.05, ** P <0.01 vs. control. F , confocal images showing the membranous and intracellular distributions of Kv4.2. GFP ( green ) and anti-Flag (red ) represented the total and membranous Kv4.2, respectively. Note that MWCNT-treated cells showed reduced membranous Kv4.2 compared with control cells. Scale bars = 50 µm.

Techniques Used: Flow Cytometry, Fluorescence, Negative Control, Expressing, SDS Page, Co-Immunoprecipitation Assay


Structured Review

Abcam kchip2
Sequence of primers used for quantitative real-time PCR.
Kchip2, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/kchip2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
kchip2 - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "MicroRNA-301a Mediated Regulation of Kv4.2 in Diabetes: Identification of Key Modulators"

Article Title: MicroRNA-301a Mediated Regulation of Kv4.2 in Diabetes: Identification of Key Modulators

Journal: PLoS ONE

doi: 10.1371/journal.pone.0060545

Sequence of primers used for quantitative real-time PCR.
Figure Legend Snippet: Sequence of primers used for quantitative real-time PCR.

Techniques Used: Sequencing

Quantitative real-time PCR (qRT-PCR) in right ventricle (RV) of wild type (Wt) and diabetic (db/db) mouse hearts with potassium channels Kv1.4, 4.2, 2.1, 4.3, 1.5, 10.2, and sodium channels Scn1b and Scn5a, along with Kv channel gene chaperon KChIP2 (A), transcriptional factors such as GATA4, GATA6, Irx5, NFkB (B), and Hif1α, along with MHC-α, MHC-β and Gja1 (C). Bars represent mean (±SEM) expression in fold, n = 3 and * represents p≤0.05.
Figure Legend Snippet: Quantitative real-time PCR (qRT-PCR) in right ventricle (RV) of wild type (Wt) and diabetic (db/db) mouse hearts with potassium channels Kv1.4, 4.2, 2.1, 4.3, 1.5, 10.2, and sodium channels Scn1b and Scn5a, along with Kv channel gene chaperon KChIP2 (A), transcriptional factors such as GATA4, GATA6, Irx5, NFkB (B), and Hif1α, along with MHC-α, MHC-β and Gja1 (C). Bars represent mean (±SEM) expression in fold, n = 3 and * represents p≤0.05.

Techniques Used: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing

The qRT-PCR analysis of Kv4.2 and 1.4 in epicardium, endocardium and apex region (A), GATA4, GATA6, Irx5, NFkB (B), Kv2.1, Kv4.3, Scn1b, Scn5a, and KChIP2 (C) in epicardium of left ventricle (LV) from wild type (Wt) and diabetic (db/db) mouse hearts. Normalized fold values were expressed in bar diagrams are mean (±SEM) of n = 3 and * represents p≤0.05.
Figure Legend Snippet: The qRT-PCR analysis of Kv4.2 and 1.4 in epicardium, endocardium and apex region (A), GATA4, GATA6, Irx5, NFkB (B), Kv2.1, Kv4.3, Scn1b, Scn5a, and KChIP2 (C) in epicardium of left ventricle (LV) from wild type (Wt) and diabetic (db/db) mouse hearts. Normalized fold values were expressed in bar diagrams are mean (±SEM) of n = 3 and * represents p≤0.05.

Techniques Used: Quantitative RT-PCR

Comparative Western blot analysis of the key potassium channels along with transcription factors, chaperons and hypertrophic markers are shown (A). Band intensities for Kv4.2, 1.4, KChIP2 (B), Irx5, NFkB, MHC6 and MHC7 (C) were measured and presented as bar diagrams after normalizing with GAPDH band intensities. All the values presented here are mean (±SEM) of n = 5–6, and * represents p≤0.05.
Figure Legend Snippet: Comparative Western blot analysis of the key potassium channels along with transcription factors, chaperons and hypertrophic markers are shown (A). Band intensities for Kv4.2, 1.4, KChIP2 (B), Irx5, NFkB, MHC6 and MHC7 (C) were measured and presented as bar diagrams after normalizing with GAPDH band intensities. All the values presented here are mean (±SEM) of n = 5–6, and * represents p≤0.05.

Techniques Used: Western Blot

Protein profiling of differentially expressed genes in left ventricle of db/db group compared with wild type (Wt) controls (A). Band intensities for Kv4.2, 1.4, KChIP2 (B), Irx5, NFkB, MHC6 and MHC7 (C) were measured and presented as bar diagrams after normalizing with GAPDH band intensities. All the values presented here are mean (±SEM) of n = 5–6, and * represents p≤0.05.
Figure Legend Snippet: Protein profiling of differentially expressed genes in left ventricle of db/db group compared with wild type (Wt) controls (A). Band intensities for Kv4.2, 1.4, KChIP2 (B), Irx5, NFkB, MHC6 and MHC7 (C) were measured and presented as bar diagrams after normalizing with GAPDH band intensities. All the values presented here are mean (±SEM) of n = 5–6, and * represents p≤0.05.

Techniques Used:

The expression of miR-301 in db/db and Wt mouse right and left ventricles were assessed by Taq-man PCR (A). Rat cardiomyocytes (H9C2) were transfected with either scrambled or miR-301 inhibitor (50 nM) for different time points 24, 48 and 72 hours and miR-301a expressions were quantified using TaqMan assay (B). The bars labeled with 72‡ are the cells transfected with inhibitor (50 nM) for 72 h with additional supply of inhibitor at 48 h. The expression of Kv4.2, Kv4.3 KChIP2, Irx5, and NFkB were quantified using qRT-PCR for the cells transfected with miR-301a inhibitor or scrambled inhibitor for 24 h (C), 48 h (D) and 72 h (E). All values are normalized with housekeeping gene (U6 RNA for TaqMan and HPRT for qRT-PCR) and plotted as mean (±SEM) of n = 3–6. * represents p≤0.05, ** with p≤0.005, and *** with p≤0.0005.
Figure Legend Snippet: The expression of miR-301 in db/db and Wt mouse right and left ventricles were assessed by Taq-man PCR (A). Rat cardiomyocytes (H9C2) were transfected with either scrambled or miR-301 inhibitor (50 nM) for different time points 24, 48 and 72 hours and miR-301a expressions were quantified using TaqMan assay (B). The bars labeled with 72‡ are the cells transfected with inhibitor (50 nM) for 72 h with additional supply of inhibitor at 48 h. The expression of Kv4.2, Kv4.3 KChIP2, Irx5, and NFkB were quantified using qRT-PCR for the cells transfected with miR-301a inhibitor or scrambled inhibitor for 24 h (C), 48 h (D) and 72 h (E). All values are normalized with housekeeping gene (U6 RNA for TaqMan and HPRT for qRT-PCR) and plotted as mean (±SEM) of n = 3–6. * represents p≤0.05, ** with p≤0.005, and *** with p≤0.0005.

Techniques Used: Expressing, Transfection, TaqMan Assay, Labeling, Quantitative RT-PCR


Structured Review

Abcam anti kchip2 antibody
( A ) Flow chart of animal experiments. ( B and D ) Determinations of IS levels in serum ( B ) and heart tissues ( D ) at 8 weeks after right nephrectomy ( n = 5 per group). ( C and E ) Measurements of IS levels in serum ( C ) and heart tissues ( E ) after 8 weeks of IS treatment ( n = 6 per group). ( F and I ) Representative immunoblots ( F ) and average data ( I ) of Kv4.2, Kv4.3, and <t>KChIP2</t> proteins in vehicle and IS treatment groups ( n = 6 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, and KChIP2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group). ( J ) Representative IHC images of Kv4.2 and Kv4.3 proteins in rats. Scale bar: 100 μm. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( C , E , and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( B , D , and H ). * P < 0.05, ** P < 0.01.
Anti Kchip2 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti kchip2 antibody/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti kchip2 antibody - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "Indoxyl sulfate reduces I to,f by activating ROS/MAPK and NF- κ B signaling pathways"

Article Title: Indoxyl sulfate reduces I to,f by activating ROS/MAPK and NF- κ B signaling pathways

Journal: JCI Insight

doi: 10.1172/jci.insight.145475

( A ) Flow chart of animal experiments. ( B and D ) Determinations of IS levels in serum ( B ) and heart tissues ( D ) at 8 weeks after right nephrectomy ( n = 5 per group). ( C and E ) Measurements of IS levels in serum ( C ) and heart tissues ( E ) after 8 weeks of IS treatment ( n = 6 per group). ( F and I ) Representative immunoblots ( F ) and average data ( I ) of Kv4.2, Kv4.3, and KChIP2 proteins in vehicle and IS treatment groups ( n = 6 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, and KChIP2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group). ( J ) Representative IHC images of Kv4.2 and Kv4.3 proteins in rats. Scale bar: 100 μm. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( C , E , and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( B , D , and H ). * P < 0.05, ** P < 0.01.
Figure Legend Snippet: ( A ) Flow chart of animal experiments. ( B and D ) Determinations of IS levels in serum ( B ) and heart tissues ( D ) at 8 weeks after right nephrectomy ( n = 5 per group). ( C and E ) Measurements of IS levels in serum ( C ) and heart tissues ( E ) after 8 weeks of IS treatment ( n = 6 per group). ( F and I ) Representative immunoblots ( F ) and average data ( I ) of Kv4.2, Kv4.3, and KChIP2 proteins in vehicle and IS treatment groups ( n = 6 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, and KChIP2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group). ( J ) Representative IHC images of Kv4.2 and Kv4.3 proteins in rats. Scale bar: 100 μm. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( C , E , and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( B , D , and H ). * P < 0.05, ** P < 0.01.

Techniques Used: Western Blot

( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, and KChIP2 proteins in NRVMs treated with different concentrations of IS ( n =3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in NRVMs treated with different concentrations of IS ( n = 3 per group). ( D and E ) Representative immunofluorescence images of Kv4.2 ( D ) and KChIP2 ( E ) proteins in NRVMs. Scale bar: 50 μm. ( F and G ) Representative I to,f traces ( F ) and average I to,f densities (peak minus steady state) versus membrane potentials ( G ) in NRVMs treated with different concentrations of IS ( n = 5 per group).The inset in F shows the voltage-clamp protocol. ( H ) Average time constants (τ) of decay of I to,f at +60 mV in NRVMs ( n = 5 per group). ( I and J ) Average values of constants (k) of activation ( I ) and half-maximal voltage of activation(V 0.5 , act) ( J ) in NRVMs ( n = 5 per group). ( K ) Voltage-dependent activation curves of I to,f in NRVMs ( n = 5 per group). Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA followed by Bonferroni post hoc test. * P < 0.05 versus control, ** P < 0.01 versus control.
Figure Legend Snippet: ( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, and KChIP2 proteins in NRVMs treated with different concentrations of IS ( n =3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in NRVMs treated with different concentrations of IS ( n = 3 per group). ( D and E ) Representative immunofluorescence images of Kv4.2 ( D ) and KChIP2 ( E ) proteins in NRVMs. Scale bar: 50 μm. ( F and G ) Representative I to,f traces ( F ) and average I to,f densities (peak minus steady state) versus membrane potentials ( G ) in NRVMs treated with different concentrations of IS ( n = 5 per group).The inset in F shows the voltage-clamp protocol. ( H ) Average time constants (τ) of decay of I to,f at +60 mV in NRVMs ( n = 5 per group). ( I and J ) Average values of constants (k) of activation ( I ) and half-maximal voltage of activation(V 0.5 , act) ( J ) in NRVMs ( n = 5 per group). ( K ) Voltage-dependent activation curves of I to,f in NRVMs ( n = 5 per group). Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA followed by Bonferroni post hoc test. * P < 0.05 versus control, ** P < 0.01 versus control.

Techniques Used: Western Blot, Immunofluorescence, Activation Assay

( A and D ) Measurements of ROS productions based on DHE fluorescence in rat hearts. ( A ) Representative images of DHE immunofluorescence. Scale bar: 200 μm. ( D ) Relative ROS fluorescence intensities in sham, CKD, and CKD plus BB536 groups ( n = 5 per group), and in vehicle and IS treatment groups ( n = 6 per group). ( B and E ) Measurements of ROS productions based on flow cytometry in NRVMs treated with different concentrations of IS. ( B ) Representative flow cytometric histograms in NRVMs. ( E ) Relative ROS fluorescence intensities in NRVMs ( n = 5 per group). ( C and F ) NAC reversed ROS production induced by IS in NRVMs. ( C ) Representative flow cytometric histograms in 4 groups. ( F ) Relative ROS fluorescence intensities detected by flow cytometry in 4 groups ( n = 5 per group). ( G and H ) Representative immunoblots of NOX2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group) ( G ), and in vehicle and IS treatment groups ( n = 6 per group) ( H ). ( I ) Average immunoblots data of NOX2 proteins in the hearts of rats. ( J and M ) Representative immunoblots ( J ) and average data ( M ) of NOX2 proteins in NRVMs treated with different concentrations of IS ( n = 3 per group). ( K and N ) Representative immunoblots ( K ) and average data ( N ) of NOX2 proteins in control, DPI, IS, and IS plus DPI groups ( n = 3 per group). ( L and O ) Representative immunoblots ( L ) and average data ( O ) of NOX2 proteins in control, APO, IS, and IS plus APO groups ( n = 3 per group). ( P and Q ) Representative immunoblots ( P ) and average data ( Q ) of Kv4.2, Kv4.3, and KChIP2 proteins in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). ( R ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). NRVMs in IS and IS plus NAC groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( D and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( D – F , I , M – O , Q , and R ). * P < 0.05, ** P < 0.01.
Figure Legend Snippet: ( A and D ) Measurements of ROS productions based on DHE fluorescence in rat hearts. ( A ) Representative images of DHE immunofluorescence. Scale bar: 200 μm. ( D ) Relative ROS fluorescence intensities in sham, CKD, and CKD plus BB536 groups ( n = 5 per group), and in vehicle and IS treatment groups ( n = 6 per group). ( B and E ) Measurements of ROS productions based on flow cytometry in NRVMs treated with different concentrations of IS. ( B ) Representative flow cytometric histograms in NRVMs. ( E ) Relative ROS fluorescence intensities in NRVMs ( n = 5 per group). ( C and F ) NAC reversed ROS production induced by IS in NRVMs. ( C ) Representative flow cytometric histograms in 4 groups. ( F ) Relative ROS fluorescence intensities detected by flow cytometry in 4 groups ( n = 5 per group). ( G and H ) Representative immunoblots of NOX2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group) ( G ), and in vehicle and IS treatment groups ( n = 6 per group) ( H ). ( I ) Average immunoblots data of NOX2 proteins in the hearts of rats. ( J and M ) Representative immunoblots ( J ) and average data ( M ) of NOX2 proteins in NRVMs treated with different concentrations of IS ( n = 3 per group). ( K and N ) Representative immunoblots ( K ) and average data ( N ) of NOX2 proteins in control, DPI, IS, and IS plus DPI groups ( n = 3 per group). ( L and O ) Representative immunoblots ( L ) and average data ( O ) of NOX2 proteins in control, APO, IS, and IS plus APO groups ( n = 3 per group). ( P and Q ) Representative immunoblots ( P ) and average data ( Q ) of Kv4.2, Kv4.3, and KChIP2 proteins in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). ( R ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). NRVMs in IS and IS plus NAC groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( D and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( D – F , I , M – O , Q , and R ). * P < 0.05, ** P < 0.01.

Techniques Used: Fluorescence, Immunofluorescence, Flow Cytometry, Western Blot

( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, KChIP2, and P-p38 MAPK in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( D and E ) Representative immunoblots ( D ) and average data ( E ) of Kv4.2, Kv4.3, KChIP2, and P-p44/42 MAPK in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( F ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, KChIP2, and P-NF-κB in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). ( I ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). NRVMs in IS, IS plus SB, IS plus U0126, and IS plus BAY groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA, followed by Bonferroni post hoc test. * P < 0.05, ** P < 0.01.
Figure Legend Snippet: ( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, KChIP2, and P-p38 MAPK in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( D and E ) Representative immunoblots ( D ) and average data ( E ) of Kv4.2, Kv4.3, KChIP2, and P-p44/42 MAPK in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( F ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, KChIP2, and P-NF-κB in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). ( I ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). NRVMs in IS, IS plus SB, IS plus U0126, and IS plus BAY groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA, followed by Bonferroni post hoc test. * P < 0.05, ** P < 0.01.

Techniques Used: Western Blot


Structured Review

Abcam anti kchip2 antibody
( A ) Flow chart of animal experiments. ( B and D ) Determinations of IS levels in serum ( B ) and heart tissues ( D ) at 8 weeks after right nephrectomy ( n = 5 per group). ( C and E ) Measurements of IS levels in serum ( C ) and heart tissues ( E ) after 8 weeks of IS treatment ( n = 6 per group). ( F and I ) Representative immunoblots ( F ) and average data ( I ) of Kv4.2, Kv4.3, and <t>KChIP2</t> proteins in vehicle and IS treatment groups ( n = 6 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, and KChIP2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group). ( J ) Representative IHC images of Kv4.2 and Kv4.3 proteins in rats. Scale bar: 100 μm. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( C , E , and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( B , D , and H ). * P < 0.05, ** P < 0.01.
Anti Kchip2 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti kchip2 antibody/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti kchip2 antibody - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "Indoxyl sulfate reduces I to,f by activating ROS/MAPK and NF- κ B signaling pathways"

Article Title: Indoxyl sulfate reduces I to,f by activating ROS/MAPK and NF- κ B signaling pathways

Journal: JCI Insight

doi: 10.1172/jci.insight.145475

( A ) Flow chart of animal experiments. ( B and D ) Determinations of IS levels in serum ( B ) and heart tissues ( D ) at 8 weeks after right nephrectomy ( n = 5 per group). ( C and E ) Measurements of IS levels in serum ( C ) and heart tissues ( E ) after 8 weeks of IS treatment ( n = 6 per group). ( F and I ) Representative immunoblots ( F ) and average data ( I ) of Kv4.2, Kv4.3, and KChIP2 proteins in vehicle and IS treatment groups ( n = 6 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, and KChIP2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group). ( J ) Representative IHC images of Kv4.2 and Kv4.3 proteins in rats. Scale bar: 100 μm. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( C , E , and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( B , D , and H ). * P < 0.05, ** P < 0.01.
Figure Legend Snippet: ( A ) Flow chart of animal experiments. ( B and D ) Determinations of IS levels in serum ( B ) and heart tissues ( D ) at 8 weeks after right nephrectomy ( n = 5 per group). ( C and E ) Measurements of IS levels in serum ( C ) and heart tissues ( E ) after 8 weeks of IS treatment ( n = 6 per group). ( F and I ) Representative immunoblots ( F ) and average data ( I ) of Kv4.2, Kv4.3, and KChIP2 proteins in vehicle and IS treatment groups ( n = 6 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, and KChIP2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group). ( J ) Representative IHC images of Kv4.2 and Kv4.3 proteins in rats. Scale bar: 100 μm. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( C , E , and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( B , D , and H ). * P < 0.05, ** P < 0.01.

Techniques Used: Western Blot

( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, and KChIP2 proteins in NRVMs treated with different concentrations of IS ( n =3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in NRVMs treated with different concentrations of IS ( n = 3 per group). ( D and E ) Representative immunofluorescence images of Kv4.2 ( D ) and KChIP2 ( E ) proteins in NRVMs. Scale bar: 50 μm. ( F and G ) Representative I to,f traces ( F ) and average I to,f densities (peak minus steady state) versus membrane potentials ( G ) in NRVMs treated with different concentrations of IS ( n = 5 per group).The inset in F shows the voltage-clamp protocol. ( H ) Average time constants (τ) of decay of I to,f at +60 mV in NRVMs ( n = 5 per group). ( I and J ) Average values of constants (k) of activation ( I ) and half-maximal voltage of activation(V 0.5 , act) ( J ) in NRVMs ( n = 5 per group). ( K ) Voltage-dependent activation curves of I to,f in NRVMs ( n = 5 per group). Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA followed by Bonferroni post hoc test. * P < 0.05 versus control, ** P < 0.01 versus control.
Figure Legend Snippet: ( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, and KChIP2 proteins in NRVMs treated with different concentrations of IS ( n =3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in NRVMs treated with different concentrations of IS ( n = 3 per group). ( D and E ) Representative immunofluorescence images of Kv4.2 ( D ) and KChIP2 ( E ) proteins in NRVMs. Scale bar: 50 μm. ( F and G ) Representative I to,f traces ( F ) and average I to,f densities (peak minus steady state) versus membrane potentials ( G ) in NRVMs treated with different concentrations of IS ( n = 5 per group).The inset in F shows the voltage-clamp protocol. ( H ) Average time constants (τ) of decay of I to,f at +60 mV in NRVMs ( n = 5 per group). ( I and J ) Average values of constants (k) of activation ( I ) and half-maximal voltage of activation(V 0.5 , act) ( J ) in NRVMs ( n = 5 per group). ( K ) Voltage-dependent activation curves of I to,f in NRVMs ( n = 5 per group). Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA followed by Bonferroni post hoc test. * P < 0.05 versus control, ** P < 0.01 versus control.

Techniques Used: Western Blot, Immunofluorescence, Activation Assay

( A and D ) Measurements of ROS productions based on DHE fluorescence in rat hearts. ( A ) Representative images of DHE immunofluorescence. Scale bar: 200 μm. ( D ) Relative ROS fluorescence intensities in sham, CKD, and CKD plus BB536 groups ( n = 5 per group), and in vehicle and IS treatment groups ( n = 6 per group). ( B and E ) Measurements of ROS productions based on flow cytometry in NRVMs treated with different concentrations of IS. ( B ) Representative flow cytometric histograms in NRVMs. ( E ) Relative ROS fluorescence intensities in NRVMs ( n = 5 per group). ( C and F ) NAC reversed ROS production induced by IS in NRVMs. ( C ) Representative flow cytometric histograms in 4 groups. ( F ) Relative ROS fluorescence intensities detected by flow cytometry in 4 groups ( n = 5 per group). ( G and H ) Representative immunoblots of NOX2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group) ( G ), and in vehicle and IS treatment groups ( n = 6 per group) ( H ). ( I ) Average immunoblots data of NOX2 proteins in the hearts of rats. ( J and M ) Representative immunoblots ( J ) and average data ( M ) of NOX2 proteins in NRVMs treated with different concentrations of IS ( n = 3 per group). ( K and N ) Representative immunoblots ( K ) and average data ( N ) of NOX2 proteins in control, DPI, IS, and IS plus DPI groups ( n = 3 per group). ( L and O ) Representative immunoblots ( L ) and average data ( O ) of NOX2 proteins in control, APO, IS, and IS plus APO groups ( n = 3 per group). ( P and Q ) Representative immunoblots ( P ) and average data ( Q ) of Kv4.2, Kv4.3, and KChIP2 proteins in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). ( R ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). NRVMs in IS and IS plus NAC groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( D and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( D – F , I , M – O , Q , and R ). * P < 0.05, ** P < 0.01.
Figure Legend Snippet: ( A and D ) Measurements of ROS productions based on DHE fluorescence in rat hearts. ( A ) Representative images of DHE immunofluorescence. Scale bar: 200 μm. ( D ) Relative ROS fluorescence intensities in sham, CKD, and CKD plus BB536 groups ( n = 5 per group), and in vehicle and IS treatment groups ( n = 6 per group). ( B and E ) Measurements of ROS productions based on flow cytometry in NRVMs treated with different concentrations of IS. ( B ) Representative flow cytometric histograms in NRVMs. ( E ) Relative ROS fluorescence intensities in NRVMs ( n = 5 per group). ( C and F ) NAC reversed ROS production induced by IS in NRVMs. ( C ) Representative flow cytometric histograms in 4 groups. ( F ) Relative ROS fluorescence intensities detected by flow cytometry in 4 groups ( n = 5 per group). ( G and H ) Representative immunoblots of NOX2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group) ( G ), and in vehicle and IS treatment groups ( n = 6 per group) ( H ). ( I ) Average immunoblots data of NOX2 proteins in the hearts of rats. ( J and M ) Representative immunoblots ( J ) and average data ( M ) of NOX2 proteins in NRVMs treated with different concentrations of IS ( n = 3 per group). ( K and N ) Representative immunoblots ( K ) and average data ( N ) of NOX2 proteins in control, DPI, IS, and IS plus DPI groups ( n = 3 per group). ( L and O ) Representative immunoblots ( L ) and average data ( O ) of NOX2 proteins in control, APO, IS, and IS plus APO groups ( n = 3 per group). ( P and Q ) Representative immunoblots ( P ) and average data ( Q ) of Kv4.2, Kv4.3, and KChIP2 proteins in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). ( R ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). NRVMs in IS and IS plus NAC groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( D and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( D – F , I , M – O , Q , and R ). * P < 0.05, ** P < 0.01.

Techniques Used: Fluorescence, Immunofluorescence, Flow Cytometry, Western Blot

( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, KChIP2, and P-p38 MAPK in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( D and E ) Representative immunoblots ( D ) and average data ( E ) of Kv4.2, Kv4.3, KChIP2, and P-p44/42 MAPK in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( F ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, KChIP2, and P-NF-κB in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). ( I ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). NRVMs in IS, IS plus SB, IS plus U0126, and IS plus BAY groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA, followed by Bonferroni post hoc test. * P < 0.05, ** P < 0.01.
Figure Legend Snippet: ( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, KChIP2, and P-p38 MAPK in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( D and E ) Representative immunoblots ( D ) and average data ( E ) of Kv4.2, Kv4.3, KChIP2, and P-p44/42 MAPK in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( F ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, KChIP2, and P-NF-κB in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). ( I ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). NRVMs in IS, IS plus SB, IS plus U0126, and IS plus BAY groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA, followed by Bonferroni post hoc test. * P < 0.05, ** P < 0.01.

Techniques Used: Western Blot


Structured Review

Abcam anti kchip2
(A) Differential interference contrast (DIC) and confocal images showing Kvβ2-associated fluorescence (red) in isolated cardiac myocytes from wild type (wt) and Kvβ2−/− animals. Nuclei (dapi) are shown in blue. (B) Western blots showing immunoreactive bands for Kv pore-forming and auxiliary subunits at respective predicted molecular weights as indicated in heart lysates from wt (n = 3) and Kvβ2−/− (n = 3) animals. As a representative loading control, immunoreactive bands for GAPDH are shown for each lane of Kv1.5 blot. (C) Summarized densitometric data for Kv1.4, Kv1.5, Kv2.1, Kv4.2, Kv4.3, Kvβ1.1, Kvβ1.2, Kvβ2, and <t>KChIP2</t> proteins in heart lysates of wt animals. Data are normalized to GAPDH (run for each blot) and expressed relative to wt controls. *P<0.05. (D, E) Representative Western blot images (D) showing immunoreactive bands for Kv1.4, Kv1.5, Kv4.2, and Kv4.3 and cadherin in cardiac myocyte membrane (m) and soluble (s) fractions and summary densitometric data (E) demonstrating membrane abundance of Kv1.4, Kv1.5, Kv4.2, and Kv4.3, expressed as the membrane:soluble band densities in hearts from Kvβ2−/− mice relative to that in hearts from wild type mice. (n = 3–5 hearts) *P<0.05.
Anti Kchip2, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti kchip2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti kchip2 - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "Metabolic regulation of Kv channels and cardiac repolarization by Kvβ2 subunits"

Article Title: Metabolic regulation of Kv channels and cardiac repolarization by Kvβ2 subunits

Journal: Journal of molecular and cellular cardiology

doi: 10.1016/j.yjmcc.2019.09.013

(A) Differential interference contrast (DIC) and confocal images showing Kvβ2-associated fluorescence (red) in isolated cardiac myocytes from wild type (wt) and Kvβ2−/− animals. Nuclei (dapi) are shown in blue. (B) Western blots showing immunoreactive bands for Kv pore-forming and auxiliary subunits at respective predicted molecular weights as indicated in heart lysates from wt (n = 3) and Kvβ2−/− (n = 3) animals. As a representative loading control, immunoreactive bands for GAPDH are shown for each lane of Kv1.5 blot. (C) Summarized densitometric data for Kv1.4, Kv1.5, Kv2.1, Kv4.2, Kv4.3, Kvβ1.1, Kvβ1.2, Kvβ2, and KChIP2 proteins in heart lysates of wt animals. Data are normalized to GAPDH (run for each blot) and expressed relative to wt controls. *P<0.05. (D, E) Representative Western blot images (D) showing immunoreactive bands for Kv1.4, Kv1.5, Kv4.2, and Kv4.3 and cadherin in cardiac myocyte membrane (m) and soluble (s) fractions and summary densitometric data (E) demonstrating membrane abundance of Kv1.4, Kv1.5, Kv4.2, and Kv4.3, expressed as the membrane:soluble band densities in hearts from Kvβ2−/− mice relative to that in hearts from wild type mice. (n = 3–5 hearts) *P<0.05.
Figure Legend Snippet: (A) Differential interference contrast (DIC) and confocal images showing Kvβ2-associated fluorescence (red) in isolated cardiac myocytes from wild type (wt) and Kvβ2−/− animals. Nuclei (dapi) are shown in blue. (B) Western blots showing immunoreactive bands for Kv pore-forming and auxiliary subunits at respective predicted molecular weights as indicated in heart lysates from wt (n = 3) and Kvβ2−/− (n = 3) animals. As a representative loading control, immunoreactive bands for GAPDH are shown for each lane of Kv1.5 blot. (C) Summarized densitometric data for Kv1.4, Kv1.5, Kv2.1, Kv4.2, Kv4.3, Kvβ1.1, Kvβ1.2, Kvβ2, and KChIP2 proteins in heart lysates of wt animals. Data are normalized to GAPDH (run for each blot) and expressed relative to wt controls. *P<0.05. (D, E) Representative Western blot images (D) showing immunoreactive bands for Kv1.4, Kv1.5, Kv4.2, and Kv4.3 and cadherin in cardiac myocyte membrane (m) and soluble (s) fractions and summary densitometric data (E) demonstrating membrane abundance of Kv1.4, Kv1.5, Kv4.2, and Kv4.3, expressed as the membrane:soluble band densities in hearts from Kvβ2−/− mice relative to that in hearts from wild type mice. (n = 3–5 hearts) *P<0.05.

Techniques Used: Fluorescence, Isolation, Western Blot


Structured Review

Abcam anti kchip2
(A) Differential interference contrast (DIC) and confocal images showing Kvβ2-associated fluorescence (red) in isolated cardiac myocytes from wild type (wt) and Kvβ2−/− animals. Nuclei (dapi) are shown in blue. (B) Western blots showing immunoreactive bands for Kv pore-forming and auxiliary subunits at respective predicted molecular weights as indicated in heart lysates from wt (n = 3) and Kvβ2−/− (n = 3) animals. As a representative loading control, immunoreactive bands for GAPDH are shown for each lane of Kv1.5 blot. (C) Summarized densitometric data for Kv1.4, Kv1.5, Kv2.1, Kv4.2, Kv4.3, Kvβ1.1, Kvβ1.2, Kvβ2, and <t>KChIP2</t> proteins in heart lysates of wt animals. Data are normalized to GAPDH (run for each blot) and expressed relative to wt controls. *P<0.05. (D, E) Representative Western blot images (D) showing immunoreactive bands for Kv1.4, Kv1.5, Kv4.2, and Kv4.3 and cadherin in cardiac myocyte membrane (m) and soluble (s) fractions and summary densitometric data (E) demonstrating membrane abundance of Kv1.4, Kv1.5, Kv4.2, and Kv4.3, expressed as the membrane:soluble band densities in hearts from Kvβ2−/− mice relative to that in hearts from wild type mice. (n = 3–5 hearts) *P<0.05.
Anti Kchip2, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti kchip2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti kchip2 - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "Metabolic regulation of Kv channels and cardiac repolarization by Kvβ2 subunits"

Article Title: Metabolic regulation of Kv channels and cardiac repolarization by Kvβ2 subunits

Journal: Journal of molecular and cellular cardiology

doi: 10.1016/j.yjmcc.2019.09.013

(A) Differential interference contrast (DIC) and confocal images showing Kvβ2-associated fluorescence (red) in isolated cardiac myocytes from wild type (wt) and Kvβ2−/− animals. Nuclei (dapi) are shown in blue. (B) Western blots showing immunoreactive bands for Kv pore-forming and auxiliary subunits at respective predicted molecular weights as indicated in heart lysates from wt (n = 3) and Kvβ2−/− (n = 3) animals. As a representative loading control, immunoreactive bands for GAPDH are shown for each lane of Kv1.5 blot. (C) Summarized densitometric data for Kv1.4, Kv1.5, Kv2.1, Kv4.2, Kv4.3, Kvβ1.1, Kvβ1.2, Kvβ2, and KChIP2 proteins in heart lysates of wt animals. Data are normalized to GAPDH (run for each blot) and expressed relative to wt controls. *P<0.05. (D, E) Representative Western blot images (D) showing immunoreactive bands for Kv1.4, Kv1.5, Kv4.2, and Kv4.3 and cadherin in cardiac myocyte membrane (m) and soluble (s) fractions and summary densitometric data (E) demonstrating membrane abundance of Kv1.4, Kv1.5, Kv4.2, and Kv4.3, expressed as the membrane:soluble band densities in hearts from Kvβ2−/− mice relative to that in hearts from wild type mice. (n = 3–5 hearts) *P<0.05.
Figure Legend Snippet: (A) Differential interference contrast (DIC) and confocal images showing Kvβ2-associated fluorescence (red) in isolated cardiac myocytes from wild type (wt) and Kvβ2−/− animals. Nuclei (dapi) are shown in blue. (B) Western blots showing immunoreactive bands for Kv pore-forming and auxiliary subunits at respective predicted molecular weights as indicated in heart lysates from wt (n = 3) and Kvβ2−/− (n = 3) animals. As a representative loading control, immunoreactive bands for GAPDH are shown for each lane of Kv1.5 blot. (C) Summarized densitometric data for Kv1.4, Kv1.5, Kv2.1, Kv4.2, Kv4.3, Kvβ1.1, Kvβ1.2, Kvβ2, and KChIP2 proteins in heart lysates of wt animals. Data are normalized to GAPDH (run for each blot) and expressed relative to wt controls. *P<0.05. (D, E) Representative Western blot images (D) showing immunoreactive bands for Kv1.4, Kv1.5, Kv4.2, and Kv4.3 and cadherin in cardiac myocyte membrane (m) and soluble (s) fractions and summary densitometric data (E) demonstrating membrane abundance of Kv1.4, Kv1.5, Kv4.2, and Kv4.3, expressed as the membrane:soluble band densities in hearts from Kvβ2−/− mice relative to that in hearts from wild type mice. (n = 3–5 hearts) *P<0.05.

Techniques Used: Fluorescence, Isolation, Western Blot


Structured Review

Abcam kchip2
Kchip2, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/kchip2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
kchip2 - by Bioz Stars, 2023-01
86/100 stars

Images


Structured Review

Abcam kchip2
Kchip2, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/kchip2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
kchip2 - by Bioz Stars, 2023-01
86/100 stars

Images


Structured Review

Abcam kchip2
Kchip2, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/kchip2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
kchip2 - by Bioz Stars, 2023-01
86/100 stars

Images


Structured Review

Abcam kchip2
Kchip2, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/kchip2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
kchip2 - by Bioz Stars, 2023-01
86/100 stars

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    Abcam anti kchip2
    A , diagram for Flag-Kv4.2-GFP oriented in the plasma membrane. Flag and GFP were tagged in the extracellular S1-S2 loop and the C-terminal, respectively. B , Western blotting analysis of HEK293 cells expressing Kv4.2 with or without the expression of <t>KChIP2.</t> The plasmids transfected in HEK293 cells were indicated above each lane. The detecting antibodies were indicated at the right side. Lane 1 showed the lysate from cells expressing the corresponding vector as control. Lane 2 and 3 represented cells expressing Kv4.2 alone or with KChIP2, respectively. C , an example of I Kv4.2 recorded in HEK293 cells expressing Flag-Kv4.2-GFP. No significant difference of I Kv4.2 was found between Flag-Kv4.2-GFP-expressing cells and cells expressing untagged Kv4.2 (not shown). D , confocal images of HEK293 cells transfected with Flag-Kv4.2-GFP, showing subcellular localization of Kv4.2, with (P (+)) or without (P (−)) membrane permeabilization. The surface population of Kv4.2 was visualized by anti-Flag antibody ( red ) and the whole Kv4.2 was indicated by GFP ( green ). The nuclei were stained by diamidino-phenyl-indole (DAPI) ( blue ). Note that anti-Flag antibody detected only the membranous Kv4.2 when without permeabilization, while it detected the overall Kv4.2 after permeabilization. The merged images showed perfect co-localization of Flag-Kv4.2-GFP detected by anti-Flag antibody and GFP, respectively. Scale bars = 50 µm.
    Anti Kchip2, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti kchip2/product/Abcam
    Average 86 stars, based on 1 article reviews
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    anti kchip2 - by Bioz Stars, 2023-01
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    86
    Abcam kchip2
    Sequence of primers used for quantitative real-time PCR.
    Kchip2, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/kchip2/product/Abcam
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    kchip2 - by Bioz Stars, 2023-01
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    86
    Abcam anti kchip2 antibody
    ( A ) Flow chart of animal experiments. ( B and D ) Determinations of IS levels in serum ( B ) and heart tissues ( D ) at 8 weeks after right nephrectomy ( n = 5 per group). ( C and E ) Measurements of IS levels in serum ( C ) and heart tissues ( E ) after 8 weeks of IS treatment ( n = 6 per group). ( F and I ) Representative immunoblots ( F ) and average data ( I ) of Kv4.2, Kv4.3, and <t>KChIP2</t> proteins in vehicle and IS treatment groups ( n = 6 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, and KChIP2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group). ( J ) Representative IHC images of Kv4.2 and Kv4.3 proteins in rats. Scale bar: 100 μm. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( C , E , and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( B , D , and H ). * P < 0.05, ** P < 0.01.
    Anti Kchip2 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti kchip2 antibody/product/Abcam
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti kchip2 antibody - by Bioz Stars, 2023-01
    86/100 stars
      Buy from Supplier

    Image Search Results


    A , diagram for Flag-Kv4.2-GFP oriented in the plasma membrane. Flag and GFP were tagged in the extracellular S1-S2 loop and the C-terminal, respectively. B , Western blotting analysis of HEK293 cells expressing Kv4.2 with or without the expression of KChIP2. The plasmids transfected in HEK293 cells were indicated above each lane. The detecting antibodies were indicated at the right side. Lane 1 showed the lysate from cells expressing the corresponding vector as control. Lane 2 and 3 represented cells expressing Kv4.2 alone or with KChIP2, respectively. C , an example of I Kv4.2 recorded in HEK293 cells expressing Flag-Kv4.2-GFP. No significant difference of I Kv4.2 was found between Flag-Kv4.2-GFP-expressing cells and cells expressing untagged Kv4.2 (not shown). D , confocal images of HEK293 cells transfected with Flag-Kv4.2-GFP, showing subcellular localization of Kv4.2, with (P (+)) or without (P (−)) membrane permeabilization. The surface population of Kv4.2 was visualized by anti-Flag antibody ( red ) and the whole Kv4.2 was indicated by GFP ( green ). The nuclei were stained by diamidino-phenyl-indole (DAPI) ( blue ). Note that anti-Flag antibody detected only the membranous Kv4.2 when without permeabilization, while it detected the overall Kv4.2 after permeabilization. The merged images showed perfect co-localization of Flag-Kv4.2-GFP detected by anti-Flag antibody and GFP, respectively. Scale bars = 50 µm.

    Journal: PLoS ONE

    Article Title: Multi-Walled Carbon Nanotubes Impair Kv4.2/4.3 Channel Activities, Delay Membrane Repolarization and Induce Bradyarrhythmias in the Rat

    doi: 10.1371/journal.pone.0101545

    Figure Lengend Snippet: A , diagram for Flag-Kv4.2-GFP oriented in the plasma membrane. Flag and GFP were tagged in the extracellular S1-S2 loop and the C-terminal, respectively. B , Western blotting analysis of HEK293 cells expressing Kv4.2 with or without the expression of KChIP2. The plasmids transfected in HEK293 cells were indicated above each lane. The detecting antibodies were indicated at the right side. Lane 1 showed the lysate from cells expressing the corresponding vector as control. Lane 2 and 3 represented cells expressing Kv4.2 alone or with KChIP2, respectively. C , an example of I Kv4.2 recorded in HEK293 cells expressing Flag-Kv4.2-GFP. No significant difference of I Kv4.2 was found between Flag-Kv4.2-GFP-expressing cells and cells expressing untagged Kv4.2 (not shown). D , confocal images of HEK293 cells transfected with Flag-Kv4.2-GFP, showing subcellular localization of Kv4.2, with (P (+)) or without (P (−)) membrane permeabilization. The surface population of Kv4.2 was visualized by anti-Flag antibody ( red ) and the whole Kv4.2 was indicated by GFP ( green ). The nuclei were stained by diamidino-phenyl-indole (DAPI) ( blue ). Note that anti-Flag antibody detected only the membranous Kv4.2 when without permeabilization, while it detected the overall Kv4.2 after permeabilization. The merged images showed perfect co-localization of Flag-Kv4.2-GFP detected by anti-Flag antibody and GFP, respectively. Scale bars = 50 µm.

    Article Snippet: Fifty micrograms of total proteins were separated by 10% SDS-PAGE and then transferred to PVDF membrane followed by blocking and incubation with anti-Kv4.2 (1∶1000, Abcam, UK), anti-Flag for Kv4.3 (1∶1000, Sigma, USA) or anti-KChIP2 (1∶2000, Abcam, UK) primary antibodies at 4°C overnight.

    Techniques: Western Blot, Expressing, Transfection, Plasmid Preparation, Staining

    A and B , examples of activation ( A ) and inactivation ( B ) currents from transfected HEK293 cells. C and D , Boltzmann equation-fitted activation and inactivation curves, respectively. E , I Kv4.2 obtained by depolarizing the cell from −90 mV to +50 mV, showing accelerated decay upon MWCNTs treatment in HEK293 cells co-expressing Kv4.2 with KChIP2. F , statistical data of decay time constants (τ decay ) in various conditions. Abbreviations: C, carboxylated MWCNTs; P, pristine MWCNTs; A, aminated MWCNTs. ** P <0.01 vs. Kv4.2. ## P <0.01 vs. KChIP2.

    Journal: PLoS ONE

    Article Title: Multi-Walled Carbon Nanotubes Impair Kv4.2/4.3 Channel Activities, Delay Membrane Repolarization and Induce Bradyarrhythmias in the Rat

    doi: 10.1371/journal.pone.0101545

    Figure Lengend Snippet: A and B , examples of activation ( A ) and inactivation ( B ) currents from transfected HEK293 cells. C and D , Boltzmann equation-fitted activation and inactivation curves, respectively. E , I Kv4.2 obtained by depolarizing the cell from −90 mV to +50 mV, showing accelerated decay upon MWCNTs treatment in HEK293 cells co-expressing Kv4.2 with KChIP2. F , statistical data of decay time constants (τ decay ) in various conditions. Abbreviations: C, carboxylated MWCNTs; P, pristine MWCNTs; A, aminated MWCNTs. ** P <0.01 vs. Kv4.2. ## P <0.01 vs. KChIP2.

    Article Snippet: Fifty micrograms of total proteins were separated by 10% SDS-PAGE and then transferred to PVDF membrane followed by blocking and incubation with anti-Kv4.2 (1∶1000, Abcam, UK), anti-Flag for Kv4.3 (1∶1000, Sigma, USA) or anti-KChIP2 (1∶2000, Abcam, UK) primary antibodies at 4°C overnight.

    Techniques: Activation Assay, Transfection, Expressing

    A , I-V curves of I Kv4.2 at different conditions. B , the voltage-dependent activation and inactivation curves fitted with Boltzmann equation. C and D , effects of MWCNTs on the decay kinetics of HEK293 cells expressing Kv4.2 with or without KChIP2. ** P <0.01 vs. control. E and F , the recovery curve and the statistical recovery time constant (τ recovery ), respectively. ** P <0.01 vs. control.

    Journal: PLoS ONE

    Article Title: Multi-Walled Carbon Nanotubes Impair Kv4.2/4.3 Channel Activities, Delay Membrane Repolarization and Induce Bradyarrhythmias in the Rat

    doi: 10.1371/journal.pone.0101545

    Figure Lengend Snippet: A , I-V curves of I Kv4.2 at different conditions. B , the voltage-dependent activation and inactivation curves fitted with Boltzmann equation. C and D , effects of MWCNTs on the decay kinetics of HEK293 cells expressing Kv4.2 with or without KChIP2. ** P <0.01 vs. control. E and F , the recovery curve and the statistical recovery time constant (τ recovery ), respectively. ** P <0.01 vs. control.

    Article Snippet: Fifty micrograms of total proteins were separated by 10% SDS-PAGE and then transferred to PVDF membrane followed by blocking and incubation with anti-Kv4.2 (1∶1000, Abcam, UK), anti-Flag for Kv4.3 (1∶1000, Sigma, USA) or anti-KChIP2 (1∶2000, Abcam, UK) primary antibodies at 4°C overnight.

    Techniques: Activation Assay, Expressing

    A , Western blotting analysis showing changes of Kv4.2 and KChIP2 in two cell lines. B , statistical histograms from showing the fold changes after 6 h-treatment with MWCNTs. C , biotinylation results showing the surface population of Kv4.2, with the expression of endogenous Na + /K + -ATPase (NKP) as loading control. D , quantification histograms indicating the fold changes of the surface population of Kv4.2. * P <0.05 vs . control.

    Journal: PLoS ONE

    Article Title: Multi-Walled Carbon Nanotubes Impair Kv4.2/4.3 Channel Activities, Delay Membrane Repolarization and Induce Bradyarrhythmias in the Rat

    doi: 10.1371/journal.pone.0101545

    Figure Lengend Snippet: A , Western blotting analysis showing changes of Kv4.2 and KChIP2 in two cell lines. B , statistical histograms from showing the fold changes after 6 h-treatment with MWCNTs. C , biotinylation results showing the surface population of Kv4.2, with the expression of endogenous Na + /K + -ATPase (NKP) as loading control. D , quantification histograms indicating the fold changes of the surface population of Kv4.2. * P <0.05 vs . control.

    Article Snippet: Fifty micrograms of total proteins were separated by 10% SDS-PAGE and then transferred to PVDF membrane followed by blocking and incubation with anti-Kv4.2 (1∶1000, Abcam, UK), anti-Flag for Kv4.3 (1∶1000, Sigma, USA) or anti-KChIP2 (1∶2000, Abcam, UK) primary antibodies at 4°C overnight.

    Techniques: Western Blot, Expressing

    A , an original recording of flow cytometry showing the fluorescence of Flag which represented the population of Kv4.2 in cell membrane. A leftward shift of the trace meant a reduction of fluorescence intensity in membrane and vise versa. Untransfected HEK293 cells served as negative control. B , ratio of mean fluorescence intensity of Flag to GFP in HEK293 cells expressing Kv4.2 with or without KChIP2 upon MWCNTs treatment from . C , aliquots of cell lysate from HEK293 cells expressing Kv4.2 and KChIP2 were mixed with either anti-Kv4.2 antibody or anti-KChIP2 antibody and precipitated with protein A-sepharose. Immune complexes were resolved by SDS-PAGE. The immunoprecipitating antibodies were indicated above each lane, and detecting antibodies were shown at the left side. D , effects of MWCNTs treatment on the ratio of KChIP2 to Kv4.2 with anti-Kv4.2 antibody as bait for co-IP in HEK293 cells and cell lysate. MWCNTs were applied to the culture medium and cell lysate for 6 h, respectively. E , statistical results of the effects of MWCNTs on the ratio of KChIP2 to Kv4.2 in HEK293 cells expressing Kv4.2 and KChIP2. * P <0.05, ** P <0.01 vs. control. F , confocal images showing the membranous and intracellular distributions of Kv4.2. GFP ( green ) and anti-Flag (red ) represented the total and membranous Kv4.2, respectively. Note that MWCNT-treated cells showed reduced membranous Kv4.2 compared with control cells. Scale bars = 50 µm.

    Journal: PLoS ONE

    Article Title: Multi-Walled Carbon Nanotubes Impair Kv4.2/4.3 Channel Activities, Delay Membrane Repolarization and Induce Bradyarrhythmias in the Rat

    doi: 10.1371/journal.pone.0101545

    Figure Lengend Snippet: A , an original recording of flow cytometry showing the fluorescence of Flag which represented the population of Kv4.2 in cell membrane. A leftward shift of the trace meant a reduction of fluorescence intensity in membrane and vise versa. Untransfected HEK293 cells served as negative control. B , ratio of mean fluorescence intensity of Flag to GFP in HEK293 cells expressing Kv4.2 with or without KChIP2 upon MWCNTs treatment from . C , aliquots of cell lysate from HEK293 cells expressing Kv4.2 and KChIP2 were mixed with either anti-Kv4.2 antibody or anti-KChIP2 antibody and precipitated with protein A-sepharose. Immune complexes were resolved by SDS-PAGE. The immunoprecipitating antibodies were indicated above each lane, and detecting antibodies were shown at the left side. D , effects of MWCNTs treatment on the ratio of KChIP2 to Kv4.2 with anti-Kv4.2 antibody as bait for co-IP in HEK293 cells and cell lysate. MWCNTs were applied to the culture medium and cell lysate for 6 h, respectively. E , statistical results of the effects of MWCNTs on the ratio of KChIP2 to Kv4.2 in HEK293 cells expressing Kv4.2 and KChIP2. * P <0.05, ** P <0.01 vs. control. F , confocal images showing the membranous and intracellular distributions of Kv4.2. GFP ( green ) and anti-Flag (red ) represented the total and membranous Kv4.2, respectively. Note that MWCNT-treated cells showed reduced membranous Kv4.2 compared with control cells. Scale bars = 50 µm.

    Article Snippet: Fifty micrograms of total proteins were separated by 10% SDS-PAGE and then transferred to PVDF membrane followed by blocking and incubation with anti-Kv4.2 (1∶1000, Abcam, UK), anti-Flag for Kv4.3 (1∶1000, Sigma, USA) or anti-KChIP2 (1∶2000, Abcam, UK) primary antibodies at 4°C overnight.

    Techniques: Flow Cytometry, Fluorescence, Negative Control, Expressing, SDS Page, Co-Immunoprecipitation Assay

    Sequence of primers used for quantitative real-time PCR.

    Journal: PLoS ONE

    Article Title: MicroRNA-301a Mediated Regulation of Kv4.2 in Diabetes: Identification of Key Modulators

    doi: 10.1371/journal.pone.0060545

    Figure Lengend Snippet: Sequence of primers used for quantitative real-time PCR.

    Article Snippet: Antibodies were obtained for Kv4.2 (1∶1000), Kv1.4 (1∶100), NFκB-p105/50 (1∶1000) and GAPDH (1∶10000) from Millipore (Billerica, MA, USA), KChIP2 (1∶1000), Irx5 (1∶100), myosin heavy chain-α/MHC-α (1∶200) and myosin heavy chain-β/MHC-β (1∶10) from Abcam (Cambridge, MA, USA).

    Techniques: Sequencing

    Quantitative real-time PCR (qRT-PCR) in right ventricle (RV) of wild type (Wt) and diabetic (db/db) mouse hearts with potassium channels Kv1.4, 4.2, 2.1, 4.3, 1.5, 10.2, and sodium channels Scn1b and Scn5a, along with Kv channel gene chaperon KChIP2 (A), transcriptional factors such as GATA4, GATA6, Irx5, NFkB (B), and Hif1α, along with MHC-α, MHC-β and Gja1 (C). Bars represent mean (±SEM) expression in fold, n = 3 and * represents p≤0.05.

    Journal: PLoS ONE

    Article Title: MicroRNA-301a Mediated Regulation of Kv4.2 in Diabetes: Identification of Key Modulators

    doi: 10.1371/journal.pone.0060545

    Figure Lengend Snippet: Quantitative real-time PCR (qRT-PCR) in right ventricle (RV) of wild type (Wt) and diabetic (db/db) mouse hearts with potassium channels Kv1.4, 4.2, 2.1, 4.3, 1.5, 10.2, and sodium channels Scn1b and Scn5a, along with Kv channel gene chaperon KChIP2 (A), transcriptional factors such as GATA4, GATA6, Irx5, NFkB (B), and Hif1α, along with MHC-α, MHC-β and Gja1 (C). Bars represent mean (±SEM) expression in fold, n = 3 and * represents p≤0.05.

    Article Snippet: Antibodies were obtained for Kv4.2 (1∶1000), Kv1.4 (1∶100), NFκB-p105/50 (1∶1000) and GAPDH (1∶10000) from Millipore (Billerica, MA, USA), KChIP2 (1∶1000), Irx5 (1∶100), myosin heavy chain-α/MHC-α (1∶200) and myosin heavy chain-β/MHC-β (1∶10) from Abcam (Cambridge, MA, USA).

    Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing

    The qRT-PCR analysis of Kv4.2 and 1.4 in epicardium, endocardium and apex region (A), GATA4, GATA6, Irx5, NFkB (B), Kv2.1, Kv4.3, Scn1b, Scn5a, and KChIP2 (C) in epicardium of left ventricle (LV) from wild type (Wt) and diabetic (db/db) mouse hearts. Normalized fold values were expressed in bar diagrams are mean (±SEM) of n = 3 and * represents p≤0.05.

    Journal: PLoS ONE

    Article Title: MicroRNA-301a Mediated Regulation of Kv4.2 in Diabetes: Identification of Key Modulators

    doi: 10.1371/journal.pone.0060545

    Figure Lengend Snippet: The qRT-PCR analysis of Kv4.2 and 1.4 in epicardium, endocardium and apex region (A), GATA4, GATA6, Irx5, NFkB (B), Kv2.1, Kv4.3, Scn1b, Scn5a, and KChIP2 (C) in epicardium of left ventricle (LV) from wild type (Wt) and diabetic (db/db) mouse hearts. Normalized fold values were expressed in bar diagrams are mean (±SEM) of n = 3 and * represents p≤0.05.

    Article Snippet: Antibodies were obtained for Kv4.2 (1∶1000), Kv1.4 (1∶100), NFκB-p105/50 (1∶1000) and GAPDH (1∶10000) from Millipore (Billerica, MA, USA), KChIP2 (1∶1000), Irx5 (1∶100), myosin heavy chain-α/MHC-α (1∶200) and myosin heavy chain-β/MHC-β (1∶10) from Abcam (Cambridge, MA, USA).

    Techniques: Quantitative RT-PCR

    Comparative Western blot analysis of the key potassium channels along with transcription factors, chaperons and hypertrophic markers are shown (A). Band intensities for Kv4.2, 1.4, KChIP2 (B), Irx5, NFkB, MHC6 and MHC7 (C) were measured and presented as bar diagrams after normalizing with GAPDH band intensities. All the values presented here are mean (±SEM) of n = 5–6, and * represents p≤0.05.

    Journal: PLoS ONE

    Article Title: MicroRNA-301a Mediated Regulation of Kv4.2 in Diabetes: Identification of Key Modulators

    doi: 10.1371/journal.pone.0060545

    Figure Lengend Snippet: Comparative Western blot analysis of the key potassium channels along with transcription factors, chaperons and hypertrophic markers are shown (A). Band intensities for Kv4.2, 1.4, KChIP2 (B), Irx5, NFkB, MHC6 and MHC7 (C) were measured and presented as bar diagrams after normalizing with GAPDH band intensities. All the values presented here are mean (±SEM) of n = 5–6, and * represents p≤0.05.

    Article Snippet: Antibodies were obtained for Kv4.2 (1∶1000), Kv1.4 (1∶100), NFκB-p105/50 (1∶1000) and GAPDH (1∶10000) from Millipore (Billerica, MA, USA), KChIP2 (1∶1000), Irx5 (1∶100), myosin heavy chain-α/MHC-α (1∶200) and myosin heavy chain-β/MHC-β (1∶10) from Abcam (Cambridge, MA, USA).

    Techniques: Western Blot

    Protein profiling of differentially expressed genes in left ventricle of db/db group compared with wild type (Wt) controls (A). Band intensities for Kv4.2, 1.4, KChIP2 (B), Irx5, NFkB, MHC6 and MHC7 (C) were measured and presented as bar diagrams after normalizing with GAPDH band intensities. All the values presented here are mean (±SEM) of n = 5–6, and * represents p≤0.05.

    Journal: PLoS ONE

    Article Title: MicroRNA-301a Mediated Regulation of Kv4.2 in Diabetes: Identification of Key Modulators

    doi: 10.1371/journal.pone.0060545

    Figure Lengend Snippet: Protein profiling of differentially expressed genes in left ventricle of db/db group compared with wild type (Wt) controls (A). Band intensities for Kv4.2, 1.4, KChIP2 (B), Irx5, NFkB, MHC6 and MHC7 (C) were measured and presented as bar diagrams after normalizing with GAPDH band intensities. All the values presented here are mean (±SEM) of n = 5–6, and * represents p≤0.05.

    Article Snippet: Antibodies were obtained for Kv4.2 (1∶1000), Kv1.4 (1∶100), NFκB-p105/50 (1∶1000) and GAPDH (1∶10000) from Millipore (Billerica, MA, USA), KChIP2 (1∶1000), Irx5 (1∶100), myosin heavy chain-α/MHC-α (1∶200) and myosin heavy chain-β/MHC-β (1∶10) from Abcam (Cambridge, MA, USA).

    Techniques:

    The expression of miR-301 in db/db and Wt mouse right and left ventricles were assessed by Taq-man PCR (A). Rat cardiomyocytes (H9C2) were transfected with either scrambled or miR-301 inhibitor (50 nM) for different time points 24, 48 and 72 hours and miR-301a expressions were quantified using TaqMan assay (B). The bars labeled with 72‡ are the cells transfected with inhibitor (50 nM) for 72 h with additional supply of inhibitor at 48 h. The expression of Kv4.2, Kv4.3 KChIP2, Irx5, and NFkB were quantified using qRT-PCR for the cells transfected with miR-301a inhibitor or scrambled inhibitor for 24 h (C), 48 h (D) and 72 h (E). All values are normalized with housekeeping gene (U6 RNA for TaqMan and HPRT for qRT-PCR) and plotted as mean (±SEM) of n = 3–6. * represents p≤0.05, ** with p≤0.005, and *** with p≤0.0005.

    Journal: PLoS ONE

    Article Title: MicroRNA-301a Mediated Regulation of Kv4.2 in Diabetes: Identification of Key Modulators

    doi: 10.1371/journal.pone.0060545

    Figure Lengend Snippet: The expression of miR-301 in db/db and Wt mouse right and left ventricles were assessed by Taq-man PCR (A). Rat cardiomyocytes (H9C2) were transfected with either scrambled or miR-301 inhibitor (50 nM) for different time points 24, 48 and 72 hours and miR-301a expressions were quantified using TaqMan assay (B). The bars labeled with 72‡ are the cells transfected with inhibitor (50 nM) for 72 h with additional supply of inhibitor at 48 h. The expression of Kv4.2, Kv4.3 KChIP2, Irx5, and NFkB were quantified using qRT-PCR for the cells transfected with miR-301a inhibitor or scrambled inhibitor for 24 h (C), 48 h (D) and 72 h (E). All values are normalized with housekeeping gene (U6 RNA for TaqMan and HPRT for qRT-PCR) and plotted as mean (±SEM) of n = 3–6. * represents p≤0.05, ** with p≤0.005, and *** with p≤0.0005.

    Article Snippet: Antibodies were obtained for Kv4.2 (1∶1000), Kv1.4 (1∶100), NFκB-p105/50 (1∶1000) and GAPDH (1∶10000) from Millipore (Billerica, MA, USA), KChIP2 (1∶1000), Irx5 (1∶100), myosin heavy chain-α/MHC-α (1∶200) and myosin heavy chain-β/MHC-β (1∶10) from Abcam (Cambridge, MA, USA).

    Techniques: Expressing, Transfection, TaqMan Assay, Labeling, Quantitative RT-PCR

    ( A ) Flow chart of animal experiments. ( B and D ) Determinations of IS levels in serum ( B ) and heart tissues ( D ) at 8 weeks after right nephrectomy ( n = 5 per group). ( C and E ) Measurements of IS levels in serum ( C ) and heart tissues ( E ) after 8 weeks of IS treatment ( n = 6 per group). ( F and I ) Representative immunoblots ( F ) and average data ( I ) of Kv4.2, Kv4.3, and KChIP2 proteins in vehicle and IS treatment groups ( n = 6 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, and KChIP2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group). ( J ) Representative IHC images of Kv4.2 and Kv4.3 proteins in rats. Scale bar: 100 μm. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( C , E , and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( B , D , and H ). * P < 0.05, ** P < 0.01.

    Journal: JCI Insight

    Article Title: Indoxyl sulfate reduces I to,f by activating ROS/MAPK and NF- κ B signaling pathways

    doi: 10.1172/jci.insight.145475

    Figure Lengend Snippet: ( A ) Flow chart of animal experiments. ( B and D ) Determinations of IS levels in serum ( B ) and heart tissues ( D ) at 8 weeks after right nephrectomy ( n = 5 per group). ( C and E ) Measurements of IS levels in serum ( C ) and heart tissues ( E ) after 8 weeks of IS treatment ( n = 6 per group). ( F and I ) Representative immunoblots ( F ) and average data ( I ) of Kv4.2, Kv4.3, and KChIP2 proteins in vehicle and IS treatment groups ( n = 6 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, and KChIP2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group). ( J ) Representative IHC images of Kv4.2 and Kv4.3 proteins in rats. Scale bar: 100 μm. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( C , E , and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( B , D , and H ). * P < 0.05, ** P < 0.01.

    Article Snippet: The membranes were blocked with 5% BSA solution at room temperature for 2 hours and incubated with corresponding primary antibodies including anti-GAPDH antibody (1:1000, ab8245, Cell Signaling Technology), anti-Kv4.2 antibody (1:1000, 21298-1-AP, Proteintech), anti-KCND3 (Kv4.3) antibody (1:1000, A6927, ABclonal), anti-KChIP2 antibody (1:1000, ab88542, Abcam), anti–phospho-p38 MAPK antibody (1:1000, 4511T, Cell Signaling Technology), anti–phospho-p44/42 MAPK (Erk1/2) antibody (1:2000, 4370T, Cell Signaling Technology), anti–phospho-NF-κB p65 antibody (1:1000, 3033S, Cell Signaling Technology), anti–p38 MAPK antibody (1:1000, 8690T, Cell Signaling Technology), anti–p44/42 MAPK (Erk1/2) antibody (1:1000, 4695T, Cell Signaling Technology), anti–NF-κB p65 antibody (1:1000, 8242S, Cell Signaling Technology), and NOX2 (1:5000, ab129068, Abcam) at 4°C overnight.

    Techniques: Western Blot

    ( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, and KChIP2 proteins in NRVMs treated with different concentrations of IS ( n =3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in NRVMs treated with different concentrations of IS ( n = 3 per group). ( D and E ) Representative immunofluorescence images of Kv4.2 ( D ) and KChIP2 ( E ) proteins in NRVMs. Scale bar: 50 μm. ( F and G ) Representative I to,f traces ( F ) and average I to,f densities (peak minus steady state) versus membrane potentials ( G ) in NRVMs treated with different concentrations of IS ( n = 5 per group).The inset in F shows the voltage-clamp protocol. ( H ) Average time constants (τ) of decay of I to,f at +60 mV in NRVMs ( n = 5 per group). ( I and J ) Average values of constants (k) of activation ( I ) and half-maximal voltage of activation(V 0.5 , act) ( J ) in NRVMs ( n = 5 per group). ( K ) Voltage-dependent activation curves of I to,f in NRVMs ( n = 5 per group). Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA followed by Bonferroni post hoc test. * P < 0.05 versus control, ** P < 0.01 versus control.

    Journal: JCI Insight

    Article Title: Indoxyl sulfate reduces I to,f by activating ROS/MAPK and NF- κ B signaling pathways

    doi: 10.1172/jci.insight.145475

    Figure Lengend Snippet: ( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, and KChIP2 proteins in NRVMs treated with different concentrations of IS ( n =3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in NRVMs treated with different concentrations of IS ( n = 3 per group). ( D and E ) Representative immunofluorescence images of Kv4.2 ( D ) and KChIP2 ( E ) proteins in NRVMs. Scale bar: 50 μm. ( F and G ) Representative I to,f traces ( F ) and average I to,f densities (peak minus steady state) versus membrane potentials ( G ) in NRVMs treated with different concentrations of IS ( n = 5 per group).The inset in F shows the voltage-clamp protocol. ( H ) Average time constants (τ) of decay of I to,f at +60 mV in NRVMs ( n = 5 per group). ( I and J ) Average values of constants (k) of activation ( I ) and half-maximal voltage of activation(V 0.5 , act) ( J ) in NRVMs ( n = 5 per group). ( K ) Voltage-dependent activation curves of I to,f in NRVMs ( n = 5 per group). Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA followed by Bonferroni post hoc test. * P < 0.05 versus control, ** P < 0.01 versus control.

    Article Snippet: The membranes were blocked with 5% BSA solution at room temperature for 2 hours and incubated with corresponding primary antibodies including anti-GAPDH antibody (1:1000, ab8245, Cell Signaling Technology), anti-Kv4.2 antibody (1:1000, 21298-1-AP, Proteintech), anti-KCND3 (Kv4.3) antibody (1:1000, A6927, ABclonal), anti-KChIP2 antibody (1:1000, ab88542, Abcam), anti–phospho-p38 MAPK antibody (1:1000, 4511T, Cell Signaling Technology), anti–phospho-p44/42 MAPK (Erk1/2) antibody (1:2000, 4370T, Cell Signaling Technology), anti–phospho-NF-κB p65 antibody (1:1000, 3033S, Cell Signaling Technology), anti–p38 MAPK antibody (1:1000, 8690T, Cell Signaling Technology), anti–p44/42 MAPK (Erk1/2) antibody (1:1000, 4695T, Cell Signaling Technology), anti–NF-κB p65 antibody (1:1000, 8242S, Cell Signaling Technology), and NOX2 (1:5000, ab129068, Abcam) at 4°C overnight.

    Techniques: Western Blot, Immunofluorescence, Activation Assay

    ( A and D ) Measurements of ROS productions based on DHE fluorescence in rat hearts. ( A ) Representative images of DHE immunofluorescence. Scale bar: 200 μm. ( D ) Relative ROS fluorescence intensities in sham, CKD, and CKD plus BB536 groups ( n = 5 per group), and in vehicle and IS treatment groups ( n = 6 per group). ( B and E ) Measurements of ROS productions based on flow cytometry in NRVMs treated with different concentrations of IS. ( B ) Representative flow cytometric histograms in NRVMs. ( E ) Relative ROS fluorescence intensities in NRVMs ( n = 5 per group). ( C and F ) NAC reversed ROS production induced by IS in NRVMs. ( C ) Representative flow cytometric histograms in 4 groups. ( F ) Relative ROS fluorescence intensities detected by flow cytometry in 4 groups ( n = 5 per group). ( G and H ) Representative immunoblots of NOX2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group) ( G ), and in vehicle and IS treatment groups ( n = 6 per group) ( H ). ( I ) Average immunoblots data of NOX2 proteins in the hearts of rats. ( J and M ) Representative immunoblots ( J ) and average data ( M ) of NOX2 proteins in NRVMs treated with different concentrations of IS ( n = 3 per group). ( K and N ) Representative immunoblots ( K ) and average data ( N ) of NOX2 proteins in control, DPI, IS, and IS plus DPI groups ( n = 3 per group). ( L and O ) Representative immunoblots ( L ) and average data ( O ) of NOX2 proteins in control, APO, IS, and IS plus APO groups ( n = 3 per group). ( P and Q ) Representative immunoblots ( P ) and average data ( Q ) of Kv4.2, Kv4.3, and KChIP2 proteins in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). ( R ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). NRVMs in IS and IS plus NAC groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( D and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( D – F , I , M – O , Q , and R ). * P < 0.05, ** P < 0.01.

    Journal: JCI Insight

    Article Title: Indoxyl sulfate reduces I to,f by activating ROS/MAPK and NF- κ B signaling pathways

    doi: 10.1172/jci.insight.145475

    Figure Lengend Snippet: ( A and D ) Measurements of ROS productions based on DHE fluorescence in rat hearts. ( A ) Representative images of DHE immunofluorescence. Scale bar: 200 μm. ( D ) Relative ROS fluorescence intensities in sham, CKD, and CKD plus BB536 groups ( n = 5 per group), and in vehicle and IS treatment groups ( n = 6 per group). ( B and E ) Measurements of ROS productions based on flow cytometry in NRVMs treated with different concentrations of IS. ( B ) Representative flow cytometric histograms in NRVMs. ( E ) Relative ROS fluorescence intensities in NRVMs ( n = 5 per group). ( C and F ) NAC reversed ROS production induced by IS in NRVMs. ( C ) Representative flow cytometric histograms in 4 groups. ( F ) Relative ROS fluorescence intensities detected by flow cytometry in 4 groups ( n = 5 per group). ( G and H ) Representative immunoblots of NOX2 proteins in sham, CKD, and CKD plus BB536 groups ( n = 5 per group) ( G ), and in vehicle and IS treatment groups ( n = 6 per group) ( H ). ( I ) Average immunoblots data of NOX2 proteins in the hearts of rats. ( J and M ) Representative immunoblots ( J ) and average data ( M ) of NOX2 proteins in NRVMs treated with different concentrations of IS ( n = 3 per group). ( K and N ) Representative immunoblots ( K ) and average data ( N ) of NOX2 proteins in control, DPI, IS, and IS plus DPI groups ( n = 3 per group). ( L and O ) Representative immunoblots ( L ) and average data ( O ) of NOX2 proteins in control, APO, IS, and IS plus APO groups ( n = 3 per group). ( P and Q ) Representative immunoblots ( P ) and average data ( Q ) of Kv4.2, Kv4.3, and KChIP2 proteins in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). ( R ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, NAC, IS, and IS plus NAC groups ( n = 3 per group). NRVMs in IS and IS plus NAC groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 2-tailed Student’s t test ( D and I ) and 1-way ANOVA followed by Bonferroni post hoc test ( D – F , I , M – O , Q , and R ). * P < 0.05, ** P < 0.01.

    Article Snippet: The membranes were blocked with 5% BSA solution at room temperature for 2 hours and incubated with corresponding primary antibodies including anti-GAPDH antibody (1:1000, ab8245, Cell Signaling Technology), anti-Kv4.2 antibody (1:1000, 21298-1-AP, Proteintech), anti-KCND3 (Kv4.3) antibody (1:1000, A6927, ABclonal), anti-KChIP2 antibody (1:1000, ab88542, Abcam), anti–phospho-p38 MAPK antibody (1:1000, 4511T, Cell Signaling Technology), anti–phospho-p44/42 MAPK (Erk1/2) antibody (1:2000, 4370T, Cell Signaling Technology), anti–phospho-NF-κB p65 antibody (1:1000, 3033S, Cell Signaling Technology), anti–p38 MAPK antibody (1:1000, 8690T, Cell Signaling Technology), anti–p44/42 MAPK (Erk1/2) antibody (1:1000, 4695T, Cell Signaling Technology), anti–NF-κB p65 antibody (1:1000, 8242S, Cell Signaling Technology), and NOX2 (1:5000, ab129068, Abcam) at 4°C overnight.

    Techniques: Fluorescence, Immunofluorescence, Flow Cytometry, Western Blot

    ( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, KChIP2, and P-p38 MAPK in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( D and E ) Representative immunoblots ( D ) and average data ( E ) of Kv4.2, Kv4.3, KChIP2, and P-p44/42 MAPK in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( F ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, KChIP2, and P-NF-κB in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). ( I ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). NRVMs in IS, IS plus SB, IS plus U0126, and IS plus BAY groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA, followed by Bonferroni post hoc test. * P < 0.05, ** P < 0.01.

    Journal: JCI Insight

    Article Title: Indoxyl sulfate reduces I to,f by activating ROS/MAPK and NF- κ B signaling pathways

    doi: 10.1172/jci.insight.145475

    Figure Lengend Snippet: ( A and B ) Representative immunoblots ( A ) and average data ( B ) of Kv4.2, Kv4.3, KChIP2, and P-p38 MAPK in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( C ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, SB, IS, and IS plus SB groups ( n = 3 per group). ( D and E ) Representative immunoblots ( D ) and average data ( E ) of Kv4.2, Kv4.3, KChIP2, and P-p44/42 MAPK in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( F ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, U0126, IS, and IS plus U0126 groups ( n = 3 per group). ( G and H ) Representative immunoblots ( G ) and average data ( H ) of Kv4.2, Kv4.3, KChIP2, and P-NF-κB in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). ( I ) Relative mRNA expressions for Kv4.2, Kv4.3, and KChIP2 in control, BAY, IS, and IS plus BAY groups ( n = 3 per group). NRVMs in IS, IS plus SB, IS plus U0126, and IS plus BAY groups were treated with 10 μM IS. Data are presented as mean ± SEM. Statistical analysis was performed using 1-way ANOVA, followed by Bonferroni post hoc test. * P < 0.05, ** P < 0.01.

    Article Snippet: The membranes were blocked with 5% BSA solution at room temperature for 2 hours and incubated with corresponding primary antibodies including anti-GAPDH antibody (1:1000, ab8245, Cell Signaling Technology), anti-Kv4.2 antibody (1:1000, 21298-1-AP, Proteintech), anti-KCND3 (Kv4.3) antibody (1:1000, A6927, ABclonal), anti-KChIP2 antibody (1:1000, ab88542, Abcam), anti–phospho-p38 MAPK antibody (1:1000, 4511T, Cell Signaling Technology), anti–phospho-p44/42 MAPK (Erk1/2) antibody (1:2000, 4370T, Cell Signaling Technology), anti–phospho-NF-κB p65 antibody (1:1000, 3033S, Cell Signaling Technology), anti–p38 MAPK antibody (1:1000, 8690T, Cell Signaling Technology), anti–p44/42 MAPK (Erk1/2) antibody (1:1000, 4695T, Cell Signaling Technology), anti–NF-κB p65 antibody (1:1000, 8242S, Cell Signaling Technology), and NOX2 (1:5000, ab129068, Abcam) at 4°C overnight.

    Techniques: Western Blot