Talraico et al. 2014 Cardiac Transcript Data

This transcriptome dataset includes the differential analysis of cardiac transcripts modulated in vivo within one hour of treatment with either isoproterenol (beta adrenergic receptor agonist, 1 mg/kg), gefitinib (epidermal growth factor inhibitor, 5 mg/kg) or both. Analyses are gefitinib versus vehicle, isoproterenol versus vehicle, gefitinib+isoproterenol versus vehicle and gefitinib+isoproterenol versus isoproterenol

Network analyses of Gef-sensitive ISO-regulated transcript groups.

<p>Pie chart depicting the proportion of ISO-regulated cardiac transcripts (adjusted for multiple testing, q <0.05) that are sensitive to Gef, as well as their Gef-sensitivity classification. Ingenuity network analysis of each group’s top associated network function revealed a common inclusion of ERK1/2 and Akt (red circles) within each network (green = group 1, red = group 2, blue = group 3, orange = group 4). Solid lines indicate a direct relationship and dotted lines indicate an indirect relationship. Arrowheads indicate activation, perpendicular tags indicate inhibition and lack of either indicates an interaction.</p

Fold-change summary of the 20 most highly altered cardiac transcripts within each treatment category.

<p>Fold-change summary of the 20 most highly altered cardiac transcripts within each treatment category.</p

Impact of Gef on βAR-mediated regulation of cardiac transcript expression.

<p>Stimulation of βAR leads to activation of G protein signaling to induce changes in transcript expression via classical cAMP-PKA-mediated signaling. βAR stimulation also induces the transactivation of EGFR, which alters transcript expression directly, or indirectly by antagonizing the classical βAR-cAMP-PKA pathway, effects that are blocked with Gef treatment.</p

Classification of ISO and Gef-mediated effects on cardiac transcript expression.

<p>A) Biovenn-generated diagram depicting the number of cardiac transcripts significantly regulated by each treatment and adjusted for multiple testing (q <0.05). Numbers 1–7 correspond to the differential transcript expression groups in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099195#pone-0099195-g001" target="_blank">Fig. 1C</a>, with numbers of transcripts in each group in parentheses. B) Ingenuity analysis revealed the ten most significantly associated molecular and cellular functions of the ISO-regulated transcripts (corresponding to differential expression groups 5 and 6 from the Biovenn diagram in A) along with the amount of Gef-regulated (groups 3 and 4) and Gef+ISO-regulated (group 7) transcripts in the same functional categories.</p

Comparison of absolute fold-change differences in cardiac transcript expression via transcriptome and RT-qPCR analyses.

<p>Comparison of absolute fold-change differences in cardiac transcript expression via transcriptome and RT-qPCR analyses.</p

Predicted network relationships between ERK1/2 and Akt and transcription regulators in response to Gef-sensitive βAR signaling.

<p>ERK1/2 and Akt (green nodes) have been shown to directly modulate the activity and/or expression of numerous proteins that themselves have direct effects on the transcription regulators HOXA9 and HNF4A (blue nodes, predicted to be inhibited in the Gef+ISO condition) and TRIM24 (red node, predicted to be activated in the Gef+ISO condition). Solid lines indicate a direct relationship and dotted lines indicate an indirect relationship. Arrowheads indicate activation, perpendicular tags indicate inhibition and lack of either indicates an interaction.</p

ISO and Gef differentially regulate acute changes in transcript expression in the mouse heart.

<p>A) C57BL/6 mice were given i.p. injections of ISO ± Gef for 10 min. Heart lysates underwent immunoblotting analysis for P-ERK1/2 and T-ERK1/2. ISO induced a 3-fold increase in P-ERK1/2 levels that was blocked by Gef pretreatment. Mean with SEM, ANOVA P value = 0.0123; *P<0.05, **P<0.01. B) RT-qPCR analysis of changes in the expression of the cAMP-regulated <i>Nr4a1</i> gene revealed a ∼20-fold increase in expression in response to ISO that was not blocked by Gef pretreatment. Data are presented as RQ with RQmin and RQmax as error bars; ΔCT ANOVA P value <0.0001; ***P<0.001, ns = not significant (P>0.05). C) Heatmap depicting significant (P<0.05) changes in cardiac transcript expression between vehicle-treated, Gef-treated, ISO-treated and Gef+ISO-treated mouse hearts as detected by transcriptome analysis. Differential transcript expression groups are listed to the right. D) Breakdown of cardiac transcripts into low, common and high abundance copies per cell categories with a comparison of the absolute number of transcripts (top bar) and relative abundance of transcripts (bottom bar).</p

RT-qPCR validation of changes in expression of Gef-sensitive ISO-regulated transcripts.

<p>RT-qPCR analysis of changes in expression of transcriptss identified via transcriptome analysis: A) <i>Sema3f</i> (ΔCT ANOVA P value = 0.0002), B) <i>Thbs1</i> (ΔCT ANOVA P value <0.0001), C) <i>Aplnr</i> (ΔCT ANOVA P value <0.0001), D) <i>Gck</i> (ΔCT ANOVA P value = 0.0029), E) <i>Mb</i> (ΔCT ANOVA P value <0.0001) and F) <i>Tef</i> (ΔCT ANOVA P value = 0.006). Data are presented as RQ with RQmin and RQmax as error bars; *P<0.05, **P<0.01, ***P<0.001, ns = not significant (P>0.05).</p

Grisanti et al. 2014 Cardiac Transcript Data

This file contains whole transcriptome data generated from the hearts of C57BL6/J mice that received infusion of the beta adrenergic receptor agonist isoproterenol (Iso; 3mg/kg/day) for 1 or 2 weeks. Sheet 1 contains the 1 week Iso versus vehicle data and sheet 2 contains the 2 week Iso versus vehicle data