KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse monoclonal anti-SOD2 | Enzo Life Sciences | Cat#ADI-SOD-110-F;RRID:AB_2750987 |
| Rabbit polyclonal anti-CHIP (Stub1) | GeneTex | Cat#GTX55810;RRID:AB_2750988 |
| Rabbit polyclonal anti-PP2A-Cα/β | Santa Cruz | Cat# sc-14020;RRID:AB_2169625 |
| Rabbit polyclonal anti-PP2Cα/β | Santa Cruz | Cat#sc-48829;RRID:AB_2169776 |
| Mouse monoclonal anti-Akt1 | GeneTex | Cat#GTX49160;RRID:AB_2750991 |
| Rabbit polyclonal anti-Akt2 | GeneTex | Cat#GTX128457;RRID:AB_2750992 |
| Goat polyclonal anti-Akt3 | Santa Cruz | Cat#11520;RRID:AB_2750993 |
| Rabbit monoclonal anti-phosp-Akt1T308 | GeneTex | Cat#GTX79150;RRID:AB_11163439 |
| Rabbit polyclonal anti-phospho-Akt1S473 | Cell signaling | Cat#4060P;RRID:AB_2750995 |
| Mouse monoclonal anti-HspA1A | GeneTex | Cat#GTX110079;RRID:AB_11171898 |
| Rabbit polyclonal anti-phospho- and total ERK1/2 | Cell Signaling | Cat#4696;RRID:AB_390780 |
| Rabbit monoclonal anti-phospho-and p38MAPK | Cell signaling | Cat#9790;RRID:AB_10831196 |
| Mouse monoclonal anti-phospho-Hsp70 Serine 631/633 | 21st Century, Boston | Custom:RRID:AB_2750999 |
| Bacterial and Virus Strains | ||
| Adenovirus-Human SOD2 | Blood Research Institute (BRI), WI | NA |
| Lentivirus-Human CHIP | BRI | NA |
| Lentivirus-Human Akt1 | BRI | NA |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Human recombinant SOD2 | Abcam | Cat#Ab84639 |
| Human recombinant Akt1 | My Bio Source | Cat#MBS203190 |
| Human recombinant (His-tagged) HspA1A | Sigma-Aldrich | Cat#SRP5190 |
| PDGF-BB | Sigma-Aldrich | Cat#P8147-1VL |
| DEPMPO-biotin | Cayman | Cat#0414736-11 |
| Mito-TEMPO | Cayman | Cat#0486522-4 |
| MitoB | Cayman | Cat#04865934-11 |
| MitoB-d15 | Cayman | Cat#0491693-3 |
| MitoP-d15 | Cayman | Cat#0486828-1 |
| MitoP | Cayman | Cat#0486829-1 |
| Sanguinarine chloride (PP2C inhibitor) | R and D systems | Cat#2302 |
| BMS 265246 | R and D systems | Cat#5654 |
| Roscovitine (Seliciclib, CYC202, CDK inhibitor) | Selleck hem | Cat#S1153 |
| Calyculin A (PP1 and PP2a inhibitor) | Sigma-Aldrich | Cat#C5552 |
| SB 202190 (p38MAPKinase inhibitor) | Sigma-Aldrich | Cat#S7067 |
| PD-098,059 (MAPK/ERK inhibitor) | Sigma-Aldrich | Cat#P215 |
| Wortmannin (PI3K inhibitor) | Cell Signaling | Cat#9901 |
| Acetylcholine | Sigma | Cat#A6625-25G |
| Hydrogen peroxide | EMD | Cat#HX0635-1 |
| MITOSOX RED | Life Technologies | Cat#M36008 |
| Potassium cyanide | Sigma | Cat#60178-100G |
| YGRKKRRQRRR-GSG (custom) | Biomatik | #SP0005-95-01199 |
| YGRKKRRQRRR-GAG (custom) | Biomatik | #SP0005-95-01199 |
| Critical Commercial Assays | ||
| Ubiquitin Enrichment Kit | ThermoFisher | Cat#89899 |
| SOD2 dismutase kit | Cayman | Cat#706002 |
| Mitochondrial isolation kit | Life Technologies | Cat#89874 |
| Subcellular protein fractionation kit | ThermoFisher | Cat#78840 |
| TNT® T7Quick Coupled T/T System | Promega | Cat#L1171 |
| FluoroTecGreenLys in vitro Translation labeling system | Promega | Cat#L5001 |
| Pierce anti-GST coated clear strip plates | ThermoFisher | Cat#151144 |
| Deposited Data | ||
| Mass spectrometry raw data | This paper | PRIDE: PXD011409 |
| Mass spectrometry phosphorylation data | This paper | PRIDE: PXD011422 |
| Experimental Models: Cell Lines | ||
| HEK293T cells | Konduri Lab (Gift) | NA |
| Pulmonary artery endothelial cells | Konduri Lab | NA |
| Experimental Models: Organisms/Strains | ||
| Akt1 Homozygous floxed mice (Female) | The Jackson Lab | Cat#JAX026474 |
| Akt1 Homozygous floxed mice (Male) | The Jackson Lab | Cat#JAX026474 |
| Sprague Dawley rat female | Envigo | 2830671 |
| Ductal ligation model of PPHN | Konduri Lab | NA |
| Oligonucleotides | ||
| forward 5-GGAACCGTAGGT GTTCGAGG-3′(CHIP) | QIAGEN | Cat#GS207 |
| Reverse 5-GTTCTGAGGGA GGGTGAAGC-3′(CHIP) | QIAGEN | Cat#SI04364136 |
| forward 5′-TTCATCAAGCCCCGCC TTT-3′(Hsp70) | Sigma | #3021492665-30/1 |
| Reverse 5′-ACTCCAGTTTCCTCCAGCGCGA-3′ (Hsp70 | Sigma | 3021492665-40/0 |
| Recombinant DNA | ||
| pBabe puroL myr Akt K179M T308A S473A | Addgene | Cat#1072 |
| pBabe puroL MYR Akt T308A, S473A | Addgene | Cat#9014 |
| pET24a Akt1 | TOPGene Tech. Inc. | NA |
| pET15b-SOD2 for in vitro transcription and translation | TOPgene Tech. Inc. | NA |
| pcDNA3.1-GST-Hsp70 | TOPgene Tech. Inc. | NA |
| pcDNA3.1-GST-Hsp70 mutants (1-400, 396-537 & 537-641) | TOPgene Tech. Inc. | NA |
| pcDNA3.1-HA-Akt1 mutants (1-150, 150-409, & 409- | TOPgene Tech Inc. | NA |
| pET21-Hsp70S631A, pET21-Hsp70S633A | TOPgene Tech Inc. | NA |
| Software and Algorithms | ||
| Ascore software | Beausoleil et al., 2006 | NA |
| YASARA software | Salomon-Ferrer et al., 2012 | NA |
| ImageJ | https://imagej.nih.gov/ij | NA |
| GraphPad Prism 7 | https://www.graphpad.com/ | NA |
| Other | ||
| Osmotic Pump | Durect Corporation | Cat#0004317 |