KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-Flag Affinity Gel | Sigma-Aldrich | Cat# A2220, RRID:AB_10063035 |
| Anti-V5 Agarose Affinity Gel | Sigma-Aldrich | Cat# A7345, RRID:AB_10062721 |
| Anti-GAPDH | Cell Signaling Technology | Cat# 3683, RRID:AB_1642205 |
| Anti-STK25 | Abcam | Cat# ab157188, RRID:AB_2725788 |
| Anti-Flag | Sigma-Aldrich | Cat# F3165, RRID:AB_259529 |
| Anti-GM130 | Cell Signaling Technology | Cat# 12480, RRID:AB_2797933 |
| Anti-PRKAR1A | Abcam | Cat# ab139695, RRID:AB_2893184 |
| Anti-pS77 PRKAR1A | Abcam | Cat#ab139682, RRID:AB_2904566 |
| Anti-pS83 PRKAR1A | Abcam | Cat#ab154851, RRID:AB_2904567 |
| Anti-PRKAR2A | Protein Tech | Cat# 10142-2-AP, RRID:AB_2922955 |
| Anti-TnI | Cell Signaling | Cat# 4002, RRID:AB_2206278 |
| Anti-TnI-pS23/S24 | Cell Signaling | Cat# 4004, RRID:AB_2206275 |
| Anti-Phospholamban | Cell Signaling Technology | Cat# 14562, RRID:AB_2798511 |
| Anti-Phospholamban-pS16/T17 | Cell Signaling Technology | Cat# 8496, RRID:AB_10949102 |
| Anti-Ryanodine Receptor 2 | Abcam | Cat#ab196355, RRID:AB_2904568 |
| Anti-Ryanodine Receptor 2-pS2808 | Abcam | Cat# ab59225, RRID:AB_946327 |
| Anti-V5 | Sigma-Aldrich | Cat# V8137, RRID:AB_261889 |
| Anti-PKA Catalytic subunit | Abcam | Cat# ab26322, RRID:AB_2170049 |
| Anti-mouse IgG (HRP conjugated) | Cell Signaling Technology | Cat# 7076, RRID:AB_330924 |
| Anti-rabbit IgG (HRP conjugated) | Cell Signaling Technology | Cat# 7074, RRID:AB_2099233 |
| Anti-CD172a/b | Biolegend | Cat# 323805; RRID:AB_830704 |
| Anti-CD90 | Invitrogen | 11-0909-42, RRID:AB_10668828 |
| Anti-TNNT2 | BD Biosciences | Cat# 565744, RRID:AB_2739341 |
| Biological samples | ||
| Human myocardial samples (IRB#AAAR0055) | National Disease Research Interchange (https://ndriresource.org/ats) | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| Y-27632 dihydrochloride | Tocris | Cat# 1254 |
| CHIR 99021 | Tocris | Cat# 4423 |
| Wnt-C59 | Tocris | Cat# 5148 |
| Forskolin | Sigma-Aldrich | Cat# F6886 |
| Lipofectamine 3000 | invitrogen | Cat# L3000015 |
| Fast SYBR Green mixture | Life Technologies | Cat# 4385612 |
| Recombinant protein STK25 | ORIGENE | Cat# TP303215 |
| Recombinant protein PRKAR1A | Abnova | Cat# H00005573_P01 |
| In vitro kinase buffer | Cell Signaling | Cat# 9802 |
| Critical commercial assays | ||
| PKA activity kit | Invitrogen | Cat# EIAPKA |
| 10-plex TMT kit | Navarrete-Perea et al. (2018) | PMID: 29734811 |
| SmartSeq Ultra Low V4 kit | Clontech/Takara | R400752 |
| Nextara XT Library Prep Kit | Illumina | FC-131-1024 |
| RNeasy Mini kit | Qiagen | Cat#74004 |
| On-column DNAse digestion | Qiagen | Cat# 79254 |
| Superscript™ III First-Strand Synthesis SuperMix | Invitrogen | Cat# 18080400 |
| Real Time Glo cell viability assay | Promega | Cat# G9712 |
| Deposited data | ||
| Sequencing of WT versus STK25KO cardiomyocytes | This paper | NCBI GEO: GSE195514 |
| Proteomic data | This paper | PRIDE ProteomeXchange: PXD031367 |
| Experimental models: Cell lines | ||
| HEK293T | ATCC | Cat# CRL-3216, RRID:CVCL_0063 |
| HiPSC (WTC cell line) | Material Transfer Agreements from Bruce Conklin, Gladstone Institute | N/A |
| Experimental models: Organisms/strains | ||
| C57BL/6J mice (AABC1503) | The Jackson Laboratory | Strain #000664 |
| Oligonucleotides | ||
| On-TargetPlus siRNA targeting STK25 | Horizon Discovery | Cat# L-004873-00-0050 |
| ON-TARGETplus non-targeting pool | Horizon Discovery | Cat# D-001810-10-50 |
| MISSION esiRNA targeting PRKAR1A | Sigma-Aldrich | Cat# EHU071341 |
| Fwd Primer for STK25: GCTCCTACCTAAAGAGCACCA | IDT | N/A |
| Rev Primer for STK25: TGGCAATGTATGTCTCCTCCAG | IDT | N/A |
| Fwd Primer for GAPDH: GGACTCATGACCACAGTCCATG | IDT | N/A |
| Rev Primer for GAPDH: CAGGGATGATGTTCTGGAGAGC | IDT | N/A |
| Recombinant DNA | ||
| CRISPR-Cas9 gRNA for STK25 knockout in iPSC | ORIGENE | Cat# KN203215G |
| CRISPR-Cas9 gRNA for STK25 knockout in mice | Synthego | Cat# sgRNA-stk25-7367, Cat# sgRNA-stk25-td10150 |
| Flag-Empty Vector control | GeneCopoeia | Cat# EX-NEG-M46 |
| Flag-WT-STK25 vector | GeneCopoeia | Cat# EX-M0142-M46 |
| Flag-K49R/T147A-STK25 vector | Vector Builder | N/A |
| V5-PRKAR1A | Vector Builder | VB200124-1141aes |
| V5-S77A/S83A-PRKAR1A | Vector Builder | VB200124-1126jtp |
| V5-S77E/S83E-PRKAR1A | Vector Builder | VB200124-1127pst |
| Software and algorithms | ||
| Real-Time Analysis Illumina | Illumina | https://www.illumina.com/informatics/sequencing-data-analysis.html |
| Bcl2fastq 2.19 | Illumina | https://support.illumina.com/sequencing/sequencing_software/bcl2fastq-conversion-software.html |
| Kallisto 0.44.0 | Bray et al. (2016) | https://pachterlab.github.io/kallisto/ |
| DESeq2 1.28.1 | Love et al. (2014) | https://bioconductor.org/packages/release/bioc/html/DESeq2.html |
| GSEA 4.10.0 | Subramanian et al. (2005) | https://www.gsea-msigdb.org/gsea/index.jsp |