Sequence of primers used for quantitative real-time PCR.

<p>Sequence of primers used for quantitative real-time PCR.</p

Quantitative real-time PCR of differentially expressed genes in right ventricle of diabetic mice.

<p>Quantitative real-time PCR of differentially expressed genes in right ventricle of diabetic mice.</p

Western analysis of ion channel expression and key proteins.

<p>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.</p

Cellular pathway affecting the cardiac arrhythmia, cardiovascular disease and tachycardia in RV of diabetic hearts.

<p>The differentially regulated genes in qRT-PCR analysis in right ventricle of db/db hearts compared to their wild type controls were taken as input data and Ingenuity Pathway Analysis (IPA) software was used to build interactive network(s) based on the relations available from its library. Genes shown in green color were down-regulated and up-regulated genes were shown in red color, where intensity of color is proportional to the fold values. Dotted arrows represent indirect relations and solid lines represent direct relations between the genes. Genes that are present in the network but not present in the input data are shown without any color.</p

Physical parameters of the db/db and age matched wild type (Wt) mice.

<p>The averaged body weights of db/db and Wt group normalized to tibia (A), heart weights normalized to tibia (B) and body weight(C), H&E staining of db/db and Wt hearts (D), cross-sectional area of the heart (E), RV thickness (F), Septal wall thickness (G) and LV thickness (H) are presented. All values presented in the bar diagram are mean (±SEM n = 15–16) with *p≤0.05.</p

Extracellular and intracellular expressions of TNF-α in diabetic hearts.

<p>Plasma and tissue homogenates were used for measurement of TNF-α by using ELISA kit. The right and the left ventricle were evaluated separately for measuring the TNF-α in a regional specific manner. The values are presented as bar diagrams of either TNFα protein expression in pg/ml of plasma (A) or pg of TNFα/µg of total protein (B). All the values presented here are mean (±SEM) of n = 5.</p

Protein profile in right ventricle of diabetic hearts.

<p>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.</p

Kv current recording in H9C2 cells treated with miR-301a inhibitor.

<p>H9C2 cells were either transfected with miR-301a inhibitor (50 nM) or scrambled inhibitor and potassium channel currents were measured by whole cell patch clamp technique after 72 h of treatment. (A) Representative whole cell currents recorded from H9C2 cells normalized to I_peak are shown at +50 mV. (B) The tau (τ) calculated from the recordings with scrambled or miR-301a inhibitor is shown as bar graph representing mean ± SEM with p≤0.05, n = 11 for scrambled and n = 22 for miR-301a inhibitor.</p

Post-transcriptional regulation of Kv4.2 by miR-301a by direct binding.

<p>The <i>in silico</i> analysis showing the direct binding of miR-301a to the ‘seed’ sequence on 3′-UTR region of Kv4.2 gene (A). Quantification of luciferase activity in H9C2 cells showing direct binding of miR-301a to the ‘seed’ sequence of Kv4.2 gene (B). Protein expression profile of Kv4.2 (24, 48, and 72 h) and Irx5 (48 h and 72 h) genes in H9C2 cells transfected with miR-301a inhibitor (50 nM) using Western blotting analysis (C). All the values presented in the bar diagram are mean (±SEM) of n = 3–6 and * represents p≤0.05.</p

Transcriptional regulation in the right ventricle of diabetic (db/db) heart.

<p>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.</p