Journal: Diagnostics

Article Title: A Pilot Study of Exploring miRNA–Protein Interaction Networks in Pancreatic Ductal Adenocarcinoma Patients: Implications for Diagnosis and Prognosis

doi: 10.3390/diagnostics15192479

Figure Lengend Snippet: The distribution of levels of the target proteins between PDAC patients and healthy individuals. ( a ) ESR1, ( b ) HCFC1, ( c ) KCNA1, ( d ) CACNG3, and ( e ) EPC1. Differences between the groups were analyzed using the Mann–Whitney U test. Bars represent mean protein concentration, and error bars indicate standard deviation (mean ± SD). * p < 0.05 was considered statistically significant.

Article Snippet: Quantitative analysis of ESR1 (Human Estrogen Receptor Alpha, ESR1 ELISA Kit, E7871Hu), HCFC1 (Human Host Cell Factor C1, HCFC1 ELISA Kit, E0307Hu), KCNA1 (Human Voltage-Gated Potassium Channel Subunit Alpha-1, KCNA1 ELISA Kit, E1757Hu), EPC1 (Human Enhancer of Polycomb Homolog 1, EPC1 ELISA Kit, E1137529Hu), and CACNG3 (Human Voltage-Dependent Calcium Channel Subunit Gamma-3, CACNG3 ELISA Kit, E4572Hu) proteins in the serum samples of PDAC patients was performed using sandwich-format ELISA kits (BT Lab, Bioassay Technology Laboratory, Shanghai, China).

Techniques: MANN-WHITNEY, Protein Concentration, Standard Deviation

Journal: Diagnostics

Article Title: A Pilot Study of Exploring miRNA–Protein Interaction Networks in Pancreatic Ductal Adenocarcinoma Patients: Implications for Diagnosis and Prognosis

doi: 10.3390/diagnostics15192479

Figure Lengend Snippet: Scatter plot matrix demonstrating the pairwise correlations among protein serum concentration levels of ESR1, HCFC1, KCNA1, CACNG3, and EPC1. The upper triangle of the matrix displays Pearson correlation coefficients (r) with corresponding p -values. Significant positive correlations were observed for each miRNA. * p < 0.05 and ** p < 0.001 were considered statistically significant.

Article Snippet: Quantitative analysis of ESR1 (Human Estrogen Receptor Alpha, ESR1 ELISA Kit, E7871Hu), HCFC1 (Human Host Cell Factor C1, HCFC1 ELISA Kit, E0307Hu), KCNA1 (Human Voltage-Gated Potassium Channel Subunit Alpha-1, KCNA1 ELISA Kit, E1757Hu), EPC1 (Human Enhancer of Polycomb Homolog 1, EPC1 ELISA Kit, E1137529Hu), and CACNG3 (Human Voltage-Dependent Calcium Channel Subunit Gamma-3, CACNG3 ELISA Kit, E4572Hu) proteins in the serum samples of PDAC patients was performed using sandwich-format ELISA kits (BT Lab, Bioassay Technology Laboratory, Shanghai, China).

Techniques: Concentration Assay

Journal: Diagnostics

Article Title: A Pilot Study of Exploring miRNA–Protein Interaction Networks in Pancreatic Ductal Adenocarcinoma Patients: Implications for Diagnosis and Prognosis

doi: 10.3390/diagnostics15192479

Figure Lengend Snippet: Kaplan–Meier survival curves showing overall survival durations of patients based on the expression levels of target proteins. ( a ) ESR1, ( b ) HCFC1, ( c ) KCNA1, ( d ) CACNG3, and ( e ) EPC1.

Article Snippet: Quantitative analysis of ESR1 (Human Estrogen Receptor Alpha, ESR1 ELISA Kit, E7871Hu), HCFC1 (Human Host Cell Factor C1, HCFC1 ELISA Kit, E0307Hu), KCNA1 (Human Voltage-Gated Potassium Channel Subunit Alpha-1, KCNA1 ELISA Kit, E1757Hu), EPC1 (Human Enhancer of Polycomb Homolog 1, EPC1 ELISA Kit, E1137529Hu), and CACNG3 (Human Voltage-Dependent Calcium Channel Subunit Gamma-3, CACNG3 ELISA Kit, E4572Hu) proteins in the serum samples of PDAC patients was performed using sandwich-format ELISA kits (BT Lab, Bioassay Technology Laboratory, Shanghai, China).

Techniques: Expressing

Journal: Pflugers Archiv

Article Title: Interactions between the C-terminus of Kv1.5 and Kv? regulate pyridine nucleotide-dependent changes in channel gating

doi: 10.1007/s00424-012-1093-z

Figure Lengend Snippet: COS-7 cells were transfected with Kv1.5WT or KvΔC coexpressed with Kvβ2. (a, b) The voltage-dependence of activation was determined by normalizing outward currents at indicated voltages to +50mV. Kv currents were recorded with either control internal solution or solution containing NADPH or NADP+ in the patch pipette. (c) The Vh of activation measured by the deactivating tail currents is plotted for different groups; see Material and Methods for details. *p<0.05 compared with none within each group, †p<0.05 compared with WT (none) or ΔC (none) within each group.

Article Snippet: Antibodies were obtained from the following sources: pan-Kvβ and Kv1.5 N-terminus (Santa-Cruz Biotechnologies), Kv1.5 C-terminus (Alomone Labs), Kvβ2, β1.1, β1.2 (Neuromab), GST (Novagen).

Techniques: Transfection, Activation Assay, Transferring

Journal: Pflugers Archiv

Article Title: Interactions between the C-terminus of Kv1.5 and Kv? regulate pyridine nucleotide-dependent changes in channel gating

doi: 10.1007/s00424-012-1093-z

Figure Lengend Snippet: (a) Western blots of Kvβ2 (upper panel) and Kvβ3 (middle panel) pulled down by the GST-Kv1.5 C-terminus fusion peptides. Fusion proteins containing 60, 38, or 19 terminal amino acid peptides from Kvα1.5 C-terminus attached to GST or GST with unrelated peptide (Control; 30μ g each) were incubated with lysate of Kvβ2 or Kvβ3 -expressing E.coli (350 μ g total protein). Protein complexes were pulled down using GST·Bind beads, washed and eluted with 10mM glutathione. The eluate was separated by SDS-PAGE and probed with anti-pan-Kvβ antibody, an antibody directed against the C-terminus of Kv1.5 (bait) or GST; (b) Densitometric analysis of the bands in panel a. The density of the Kvβ band precipitated with GST-C60 was assigned a 100% value. †, P< 0.05 versus Control peptide (n=3-5). (c) Determination of the binding affinity of Kvβ:nucleotide complexes to the C-terminal peptide of Kv1.5. A fixed concentration of GST-C60 (30 μ g/ml) was mixed with variable concentrations of Kvβ2 in a binary complex with NADPH (Kvβ2:NADPH), or NADP+ (Kvβ2:NADP+), or none (apo-Kvβ2) and GST pull down assay was performed. Eluted protein was separated by SDS-PAGE and the gels were silver stained. Band intensities normalized to GST-C60 input were plotted as a function of Kvβ2 concentration. The experiment was repeated 6 times; data points on the graph represent average and standard error, and the lines represent the best fit to experimental data using the Hill equation. Apo:Kvβ2 did not bind GST-C60 and no measurable intensities were found by silver stain. Inset: silver stained gels showing Kvβ2 in complex with NADPH, NADP+ or without nucleotide, pulled down with GST-C60; M, marker, 37 kDa band is shown. (d) Western blots of GST beads incubated with brain lysates. The GST-C60 construct or scrambled construct was used to pull down proteins from mouse brain extract. The eluate was separated on SDS PAGE and protein bands were visualized with antibodies against the indicated Kvβ isoform, Kv1.5 C-terminus, or GST.

Article Snippet: Antibodies were obtained from the following sources: pan-Kvβ and Kv1.5 N-terminus (Santa-Cruz Biotechnologies), Kv1.5 C-terminus (Alomone Labs), Kvβ2, β1.1, β1.2 (Neuromab), GST (Novagen).

Techniques: Western Blot, Incubation, Expressing, SDS Page, Binding Assay, Concentration Assay, Pull Down Assay, Staining, Silver Staining, Marker, Construct

Journal: eLife

Article Title: Cardiac electrophysiological remodeling associated with enhanced arrhythmia susceptibility in a canine model of elite exercise

doi: 10.7554/eLife.80710

Figure Lengend Snippet: ( A and B ) Relative protein expression of Kv4.3 and KChiP2 subunits determined by western blotting in left ventricular samples of SED (n=12) and TRN (n=12) dogs, respectively. ( C ) Representative image of Kv4.3 and KChIP2 bands and their corresponding loading controls (GAPDH). ( D and E ) Relative densities of Kv4.3 and KChiP2 protein immunolabeling obtained from SED (n=30 cells/6 dogs) and TRN (n=30 cells/6 dogs) cardiomyocytes. ( F ) Representative immunofluorescence images of canine cardiomyocytes with Kv4.3 and KChiP2 immunolabeling. The ‘n’ numbers refer to the number of dogs ( A and B ) or the number of cells followed by the number of dogs from which the cells were obtained ( D and E ). Data are expressed as mean ± SEM. Blue dots represent individual data. Figure 6—source data 1. Relative protein expression of Kv4.3 subunit determined by western blotting in sedentary and trained dogs. Figure 6—source data 2. Relative protein expression of KChiP2 subunit determined by western blotting in sedentary and trained dogs. Figure 6—source data 3. Relative density of Kv4.3 subunit determined by immunocytochemistry in sedentary and trained dogs. Figure 6—source data 4. Relative density of KChiP2 subunit determined by immunocytochemistry in sedentary and trained dogs. Figure 6—source data 5. Original unedited membranes of western blots with the relevant bands clearly labeled. Figure 6—source data 6. Original files of the full raw unedited membranes of western blots.

Article Snippet: The membrane was blocked with 2.5% non-fat milk for 1 hr at room temperature and immunolabeled overnight at 4°C with anti-KChIP2 (Alomone, #APC-142, RRID: AB_2756744 ) and anti-Kv4.3 (Alomone, #APC-017, RRID: AB_2040178 ) primary antibodies diluted 1:1000.

Techniques: Expressing, Western Blot, Immunolabeling, Immunofluorescence, Immunocytochemistry, Labeling

Journal: Acta pharmacologica Sinica

Article Title: Knockout of interleukin-17A diminishes ventricular arrhythmia susceptibility in diabetic mice via inhibiting NF-κB-mediated electrical remodeling.

doi: 10.1038/s41401-021-00659-8

Figure Lengend Snippet: Fig. 4 Effects of IL-17 knockout on the Ito and KChIP2 expression in the hearts of diabetic mice. a Representative traces of the Ito. b Current density-voltage (I–V) relationship of the Ito. n = 8–11 cells. c Current density-voltage (I–V) relationship of the Iss. n = 9–16 cells. d Mean membrane capacitance of the Ito and Iss n = 9–16 cells. e The protein and mRNA levels of Kv4.2. n = 7. f The protein and mRNA levels of Kv4.3. n = 7. g The protein and mRNA levels of KChIP2. n = 5. *P < 0.05 vs. WT mice; #P < 0.05 vs. WT+DM mice. WT wild-type, IL-17 KO IL-17 knockout, WT+DM wild-type+diabetes mellitus, IL-17 KO+DM IL-17 knockout+diabetes mellitus.

Article Snippet: The primary antibodies included rabbit anti-mouse Nav1.5 (Alomone Labs, Israel), Kv4.2 (Alomone Labs, Israel), Kv4.3 (Alomone Labs, Israel), KChIP2 (Boster, China), Cav1.2 (Alomone Labs, Israel), and NF-κB (CST, USA). β-Actin was used as an internal control.

Techniques: Knock-Out, Expressing, Membrane

Journal: Acta pharmacologica Sinica

Article Title: Knockout of interleukin-17A diminishes ventricular arrhythmia susceptibility in diabetic mice via inhibiting NF-κB-mediated electrical remodeling.

doi: 10.1038/s41401-021-00659-8

Figure Lengend Snippet: Fig. 8 Knockout of IL-17 protects against ventricular arrhythmias in STZ-induced diabetic mice. Knockout of IL-17 downregulates the expression of NF-κB, which suppresses the expression of KCNIP2, which encodes potassium voltage-gated channel interacting protein 2; CACNA1C, which encodes the pore-forming subunit of the voltage-gated L-type calcium channel Cav1.2; and SCN5A, which encodes the pore-forming subunit of the voltage-gated sodium channel Nav1.5. Decreased expression of KChIP2 and Nav1.5 prolonged the APD and slowed the conduction velocity, which increased susceptibility to ventricular arrhythmias. KCNIP2, potassium voltage-gated channel interacting protein 2; CACNA1C, calcium voltage-gated channel subunit alpha 1 C; KCND2, potassium voltage-gated channel subfamily D member 2; KCND3, potassium voltage-gated channel subfamily D member 3; SCN5A, sodium voltage-gated channel alpha subunit 5; WT+DM, wild-type+diabetes mellitus; IL-17 KO+DM, IL-17 knockout+diabetes mellitus.

Article Snippet: The primary antibodies included rabbit anti-mouse Nav1.5 (Alomone Labs, Israel), Kv4.2 (Alomone Labs, Israel), Kv4.3 (Alomone Labs, Israel), KChIP2 (Boster, China), Cav1.2 (Alomone Labs, Israel), and NF-κB (CST, USA). β-Actin was used as an internal control.

Techniques: Knock-Out, Expressing