Journal: Cell reports
Article Title: Type I interferon governs immunometabolic checkpoints that coordinate inflammation during Staphylococcal infection.
doi: 10.1016/j.celrep.2024.114607
Figure Lengend Snippet: Figure 4. Nitric oxide biases macrophage cytokine production toward type I IFN during MRSA infection (A) ELISA analysis of secreted IL-1b, IFN-b, and TNF-a from mock- or 24-h MRSA-infected WT and Nos2/ BMDMs. (B) ELISA analysis of secreted IL-1b, IFN-b, and TNF-a from WT BMDM treated ± NO, donor DETA- NONOate (500 mM) for 24 h, washed with fresh media, and then mock or MRSA infected for 24 h. (C) ELISA analysis of secreted IL-1b and IFN-b after 6- or 24-h MRSA infection ± sub-toxic doses of RC inhibitors, dimethyl malonate (complex II inhibitor; 10 mM), antimycin A (complex III inhibitor; 1 mM), and piericidin A (complex I inhibitor; 100 nM). Graphs represent mean of n = 3 biological repli- cates with SD error bars. p values were calculated using two-way ANOVA with Sidak’s post-test (A– C). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Lysostaphin Sigma Cat#L7386 Critical commercial assays Cut & Run kit Cell Signaling Cat#86652 Lactate assay Cayman Chemical Cat#600450 Greiss reagent Abcam Cat#ab234044 Mito Stress Test Agilent Cat#103015-100 LookOut Mycoplasma PCR Detection Kit Sigma Cat#MP0035 MitoTracker Green Thermo Fisher Scientific Cat#M7514 MitoTracker Deep Red Thermo Fisher Scientific Cat#M22426 Propidium iodide Bio-Rad Cat#1351101 Hoechst Thermo Fisher Scientific Cat#H3570 DAPI Thermo Fisher Scientific Cat#EN62248 PowerUP SYBR Green PCR master mix Thermo Fisher Scientific Cat#A25742 RNAeasy Plus Mini kit Qiagen Cat#74134 iScript cDNA synthesis kit Bio-Rad Cat#1708890 BioMasher II homogenizers Thermo Fisher Scientific Cat#50-136-7690 Halt protease and phosphatase inhibitor cocktail Thermo Fisher Scientific Cat#78440 RevertTM 700 Total Protein Stain for Western Blot Normalization LI-COR Cat#926-11011 TNF-a ELISA R&D Systems Cat#DY 410 IL-6 ELISA R&D Systems Cat#DY 406 IL-1b ELISA R&D Systems Cat#DY 401 IFN-b ELISA R&D Systems Cat#DY 8234-05 MPO ELISA Abcam Cat#ab155458 Bio-Rad Protein Assay (Bradford assay) Bio-Rad Cat#5000001 TRIzolTM Reagent Thermo Fisher Scientific Cat#15596026 NativePAGETM Sample Prep Kit Thermo Fisher Scientific Cat#BN2008 NativeMarkTM Unstained Protein Standard Thermo Fisher Scientific Cat#LC0725 NativePAGETM 4 to 16%, Bis-Tris, 1.0 mm, Mini Protein Gels Thermo Fisher Scientific Cat#BN1002BOX NativePAGETM Running Buffer Kit Thermo Fisher Scientific Cat#BN2007 NuPAGETM Transfer Buffer Thermo Fisher Scientific Cat#NP0006 4x Laemmli Sample Buffer Bio-Rad Cat#1610747 IGEPAL CA-630 Sigma Cat#I8896 4–20% Mini-PROTEAN TGXTM Precast Protein Gels Bio-Rad Cat#4561096 Deposited data Targeted metabolomics analysis of WT and IFNAR1 deficient murine bone marrow-derived macrophages following methicillin-resistant Staphylococcus aureus infection [Dataset] This paper https://doi.org/10.7302/h15v-j558 RNAseq analysis of Tlr2/4/9 / iBMDM and paired WT controls [Dataset] This paper GEO accession # GSE271727 Experimental models: Cell lines Tlr2/4/9 / iBMDM and paired WT controls Derived in our lab from mice obtained from the laboratory of Dr. Tod Merkel, FDA20 Ifnar1 / iBMDM and paired WT controls Derived in our lab from mice obtained from the Jackson Laboratory strain #010830 (Continued on next page) Cell Reports 43, 114607, August 27, 2024 19
Techniques: Infection, Enzyme-linked Immunosorbent Assay
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