Journal: bioRxiv

Article Title: Kv11.1 (hERG) Protein Interaction Networks Connect Endocytic Trafficking to Polygenic Influences on Cardiac Repolarization

doi: 10.64898/2026.01.27.701872

Figure Lengend Snippet: (A) Cell line generation : hiPSCs were derived from patients at upper or lower extremes of QT-PGS, plus one mid-range reference donor, and differentiated into hiPSC-CMs for proteomic analysis. (B) Global proteomics : hiPSC-CMs were lysed, digested and analyzed by label-free data-independent acquisition (DIA) mass spectrometry. (C) Affinity purification/coimmunoprecipitation coupled with mass spectrometry : Kv11.1 complexes were enriched by co-immunoprecipitation using an anti-Kv11.1 antibody or bead-only controls, followed by digestion and analysis by mass spectrometry using data-dependent acquisition (DDA). Peptides were pooled within run as indicated and analyzed to characterize Kv11.1 interactors and their quantitative differences across QT-PGS groups.

Article Snippet: This procedure was adapted from Pankow, et al. Pierce Protein A/G Magnetic Beads (ThermoFisher) containing a recombinant Protein A/G (∼50.5 kDa) that combines the IgG binding domain of both Protein A and Protein G were used to cross-link anti-Kv11.1 antibodies (Cell Signaling).

Techniques: Derivative Assay, Data-independent acquisition, Mass Spectrometry, Affinity Purification, Immunoprecipitation, Data-dependent acquisition

Journal: bioRxiv

Article Title: Kv11.1 (hERG) Protein Interaction Networks Connect Endocytic Trafficking to Polygenic Influences on Cardiac Repolarization

doi: 10.64898/2026.01.27.701872

Figure Lengend Snippet: (A) Volcano plots showing log 2 fold change versus -log 10 adjusted P value for each QT-PGS group compared to bead-only background controls. Orange dashed line indicates FDR < 0.05 (two-stage step-up with Benjamini, Krieger, and Yekutieli, n = 6-8 biological replicates per condition). Blue dots are statistically enriched Kv11.1 interactors relative to bead-only background controls. Red highlight is the KCNH2 /Kv11.1 bait. (B) Heatmaps of significantly enriched protein interactors relative to bead-only controls. Rows represent proteins; columns represent samples or group means (as indicated). (C) Heatmap of mitochondrial-function proteins (GO terms manually assigned using UniProt) showing relative abundance across groups. (D) Dot plot of log 2 fold change (sample/bead-only) for mitochondrial proteins across groups. (E) Heatmap of proteins associated with quality control and trafficking. (F) Dot plot of log 2 fold change comparing all ER quality control and trafficking across groups. Statistics for dot plots were performed using one-way ANOVA with post hoc Tukey’s multiple comparisons test. Data shown as the mean ± SEM.

Article Snippet: This procedure was adapted from Pankow, et al. Pierce Protein A/G Magnetic Beads (ThermoFisher) containing a recombinant Protein A/G (∼50.5 kDa) that combines the IgG binding domain of both Protein A and Protein G were used to cross-link anti-Kv11.1 antibodies (Cell Signaling).

Techniques: Control

Journal: bioRxiv

Article Title: Kv11.1 (hERG) Protein Interaction Networks Connect Endocytic Trafficking to Polygenic Influences on Cardiac Repolarization

doi: 10.64898/2026.01.27.701872

Figure Lengend Snippet: (A) Bubble plot of the top 30 GO terms (ranked by significance) for all 319 Kv11.1 interactors. (B) Heatmap of log2 fold changes for plasma membrane-associated proteins drawn from top GO terms. (C) Heatmap of log2 fold changes for actin filament-based movement proteins comparing QT-PGS levels. (D) Volcano plot comparing high- vs low-QT-PGS Kv11.1 interactors, showing log 2 fold changes and -log 10 adjusted P values.

Article Snippet: This procedure was adapted from Pankow, et al. Pierce Protein A/G Magnetic Beads (ThermoFisher) containing a recombinant Protein A/G (∼50.5 kDa) that combines the IgG binding domain of both Protein A and Protein G were used to cross-link anti-Kv11.1 antibodies (Cell Signaling).

Techniques: Clinical Proteomics, Membrane

Journal: bioRxiv

Article Title: Kv11.1 (hERG) Protein Interaction Networks Connect Endocytic Trafficking to Polygenic Influences on Cardiac Repolarization

doi: 10.64898/2026.01.27.701872

Figure Lengend Snippet: Schematic model summarizing Kv11.1 interactions and localization across subcellular compartments and their PGS-associated differences between low and high. ID – intercalated disk, ER – endoplasmic reticulum.

Article Snippet: This procedure was adapted from Pankow, et al. Pierce Protein A/G Magnetic Beads (ThermoFisher) containing a recombinant Protein A/G (∼50.5 kDa) that combines the IgG binding domain of both Protein A and Protein G were used to cross-link anti-Kv11.1 antibodies (Cell Signaling).

Techniques:

Journal: bioRxiv

Article Title: Kv11.1 (hERG) Protein Interaction Networks Connect Endocytic Trafficking to Polygenic Influences on Cardiac Repolarization

doi: 10.64898/2026.01.27.701872

Figure Lengend Snippet: (A) Cell line generation : hiPSCs were derived from patients at upper or lower extremes of QT-PGS, plus one mid-range reference donor, and differentiated into hiPSC-CMs for proteomic analysis. (B) Global proteomics : hiPSC-CMs were lysed, digested and analyzed by label-free data-independent acquisition (DIA) mass spectrometry. (C) Affinity purification/coimmunoprecipitation coupled with mass spectrometry : Kv11.1 complexes were enriched by co-immunoprecipitation using an anti-Kv11.1 antibody or bead-only controls, followed by digestion and analysis by mass spectrometry using data-dependent acquisition (DDA). Peptides were pooled within run as indicated and analyzed to characterize Kv11.1 interactors and their quantitative differences across QT-PGS groups.

Article Snippet: Next, 5 mg of lysate representing one biological replicate were incubated for 2 hours at 4 °C with anti-Kv11.1 antibody (Cell Signaling #12889) covalently coupled to protein A/G magnetic beads (Thermo Fisher).

Techniques: Derivative Assay, Data-independent acquisition, Mass Spectrometry, Affinity Purification, Immunoprecipitation, Data-dependent acquisition

Journal: bioRxiv

Article Title: Kv11.1 (hERG) Protein Interaction Networks Connect Endocytic Trafficking to Polygenic Influences on Cardiac Repolarization

doi: 10.64898/2026.01.27.701872

Figure Lengend Snippet: (A) Volcano plots showing log 2 fold change versus -log 10 adjusted P value for each QT-PGS group compared to bead-only background controls. Orange dashed line indicates FDR < 0.05 (two-stage step-up with Benjamini, Krieger, and Yekutieli, n = 6-8 biological replicates per condition). Blue dots are statistically enriched Kv11.1 interactors relative to bead-only background controls. Red highlight is the KCNH2 /Kv11.1 bait. (B) Heatmaps of significantly enriched protein interactors relative to bead-only controls. Rows represent proteins; columns represent samples or group means (as indicated). (C) Heatmap of mitochondrial-function proteins (GO terms manually assigned using UniProt) showing relative abundance across groups. (D) Dot plot of log 2 fold change (sample/bead-only) for mitochondrial proteins across groups. (E) Heatmap of proteins associated with quality control and trafficking. (F) Dot plot of log 2 fold change comparing all ER quality control and trafficking across groups. Statistics for dot plots were performed using one-way ANOVA with post hoc Tukey’s multiple comparisons test. Data shown as the mean ± SEM.

Article Snippet: Next, 5 mg of lysate representing one biological replicate were incubated for 2 hours at 4 °C with anti-Kv11.1 antibody (Cell Signaling #12889) covalently coupled to protein A/G magnetic beads (Thermo Fisher).

Techniques: Control

Journal: bioRxiv

Article Title: Kv11.1 (hERG) Protein Interaction Networks Connect Endocytic Trafficking to Polygenic Influences on Cardiac Repolarization

doi: 10.64898/2026.01.27.701872

Figure Lengend Snippet: (A) Bubble plot of the top 30 GO terms (ranked by significance) for all 319 Kv11.1 interactors. (B) Heatmap of log2 fold changes for plasma membrane-associated proteins drawn from top GO terms. (C) Heatmap of log2 fold changes for actin filament-based movement proteins comparing QT-PGS levels. (D) Volcano plot comparing high- vs low-QT-PGS Kv11.1 interactors, showing log 2 fold changes and -log 10 adjusted P values.

Article Snippet: Next, 5 mg of lysate representing one biological replicate were incubated for 2 hours at 4 °C with anti-Kv11.1 antibody (Cell Signaling #12889) covalently coupled to protein A/G magnetic beads (Thermo Fisher).

Techniques: Clinical Proteomics, Membrane

Journal: bioRxiv

Article Title: Kv11.1 (hERG) Protein Interaction Networks Connect Endocytic Trafficking to Polygenic Influences on Cardiac Repolarization

doi: 10.64898/2026.01.27.701872

Figure Lengend Snippet: Schematic model summarizing Kv11.1 interactions and localization across subcellular compartments and their PGS-associated differences between low and high. ID – intercalated disk, ER – endoplasmic reticulum.

Article Snippet: Next, 5 mg of lysate representing one biological replicate were incubated for 2 hours at 4 °C with anti-Kv11.1 antibody (Cell Signaling #12889) covalently coupled to protein A/G magnetic beads (Thermo Fisher).

Techniques: