KEY RESOURCES TABLE.
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| MyD88 (D80F5) Rabbit mAb | Cell Signaling Technologies | Cat. #4283 |
| beta Actin Loading Control Monoclonal Antibody (BA3R), DyLight™ 680 | Invitrogen | Catalog #MA5-15739-D680 |
| Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, DyLight™ 800 | Invitrogen | Catalog #SA5-10036 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| PS 1145 dihydrochloride | Tocris | 4569 |
| Ultrapure LPS, E. coli 0111:B4 | Invivogen | tlrl-3pelps |
| PAM2CSK4:Synthetic diacylated lipopeptide; TLR2/TLR6 agonist | Invivogen | tlrl-pm2s-1 |
| Recombinant Mouse TNF-alpha | R&D Systems | aa 80-235 |
| Recombinant Mouse IL-1 beta/IL-1F2 Protein | R&D Systems | 401ML010CF |
| SuperScript™ II Reverse Transcriptase | Invitrogen | 18064014 |
| KAPA HiFi HotStart ReadyMix | Roche | HIFI |
| Recombinant RNase Inhibitor | Takara | 2313A |
| Polydimethylsiloxane | Momentive | RTV-615 |
| Human Plasma Fibronectin Purified Protein | Millipore Sigma | FC010 |
| FluoroBrite™ DMEM | Invitrogen | A1896701 |
| Critical Commercial Assays | ||
| SuperScript™ III CellsDirect™ cDNA Synthesis Kit | Invitrogen | 18080200 |
| Cell Line Nucleofector™ Kit R | Lonza | Catalog #:VVCA-1001 |
| Csf2 PrimeTime probe assay | IDT | Mm.PT.58.10456839 |
| Csf3 PrimeTime probe assay | IDT | Mm.PT.58.43222334.g |
| Cxcl2 PrimeTime probe assay | IDT | Mm.PT.58.7603454.g |
| Cxcl3 PrimeTime probe assay | IDT | Mm.PT.58.45877295.g |
| Il23a PrimeTime probe assay | IDT | Mm.PT.58.10594618.g |
| Gapdh PrimeTime probe assay | IDT | Mm.PT.39a.1 |
| Nfkbia TaqMan probe assay | ThermoFisher Scientific | Mm00477798_m1 |
| Nfkbie TaqMan probe assay | ThermoFisher Scientific | Mm01269649_m1 |
| Nextera XT DNA Library Preparation Kit | Illumina | FC-131-1024 |
| Deposited Data | ||
| RNAseq data reported in this paper | This study | GEO: GSE193053 |
| Trace/microscopy data reported in this paper | This study | Github:https://github.com/tay-lab/Sequential_NF-kB_stim DOI: 10.5281/zenodo.6626195 |
| Experimental Models: Cell Lines | ||
| p65−/−p65-DsRedJNK-KTR NIH3T3 mouse embryonic fibroblasts | Markus Covert (Stanford) | N/A |
| Oligonucleotides | ||
| Tnfaip3 qPCR FWD:GCAGCTGGAATCTCTGAAATCT | IDT | N/A |
| Tnfaip3 qPCR REV:AGTTGTCCCATTCGTCATTCC | IDT | N/A |
| Tnfaip3 qPCR PRB:FAM/AAACAGGAC/ZEN/TTTGCTACGACACTCGG/3IABkFQ/ | IDT | N/A |
| TSO oligo: AAGCAGTGGTATCAACGCAGAGTGAATrGrGrG | IDT | N/A |
| ISPCR primer:AAGCAGTGGTATCAACGCAGAGT | IDT | N/A |
| OligodT30VN:AAGCAGTGGTATCAACGCAGAGTACTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVN | IDT | N/A |
| MyD88 gRNA template:TCGCGCTTAACGTGGGAGTG | IDT | N/A |
| Recombinant DNA | ||
| pX330-U6-Chimeric_BB-CBh-hSpCas9 | Addgene | Plasmid #42230 |
| Software and Algorithms | ||
| MATLAB R2021b | Mathworks | https://www.mathworks.com/products/matlab.html |
| RStudio Build 372 | RStudio | https://www.rstudio.com/ |
| R version 4.1.2 | R-project | https://www.r-project.org/ |
| Python 3.10 | Python | https://www.python.org/ |