Thioglycollate (TG)-elicited peritoneal myeloid cells were prepared from the Slfn4-tdT mice, as described previously6 (link). Cells were treated with 100nM Tamoxifen (Tx, dissolved in DMSO) for 24h to induce Cre recombinase activity and tdTomato expression ex vivo. The Hsa-MIR130b-3p miRNA mimic (50nM, Applied Biological Materials, MCH01270) and the mimic negative control (MCH00000) were transfected into cells using Lipofectamine LTX with PLUS reagent (Thermo Fisher). The Mmu-Mir130b-3p miRNA antisense (CAGUGCAAUGAUGAAAGGGCAU) (10nM, MSTUD0173, Sigma Aldrich), Slfn4 siRNA or the scrambled controls were transfected using Lipofectamine RNAiMAX transfection reagent (Thermo Fisher) for 48h. To induce Slfn4 expression, the cells were treated with 800U/ml recombinant IFNα (R&D, #12125-1) for 24h or at different time points.
Mstud0173
Manufactured by Merck Group
MSTUD0173 is a laboratory equipment product manufactured by Merck Group. It is a precision instrument designed for specialized laboratory applications. The core function of this product is to perform accurate measurements and analyses as required by scientific research and testing procedures. Further details about its intended use or specific applications are not available.
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2 protocols using mstud0173
1
Myeloid Cell Manipulation in Slfn4-tdT Mice
2
Inducing Slfn4 Expression in Myeloid Cells
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TG-elicited peritoneal myeloid cells were prepared from the Slfn4-tdT mice, as described previously.6 (link) Cells were treated with 100 nM Tamoxifen (Tx, dissolved in DMSO) for 24 hours to induce Cre recombinase activity and tdTomato expression ex vivo. The Hsa-MIR130b-3p miRNA mimic (50 nM, Applied Biological Materials, MCH01270) and the mimic negative control (MCH00000) were transfected into cells using Lipofectamine LTX with PLUS reagent (Thermo Fisher). The Mmu-Mir130b-3p miRNA antisense (CAGUGCAAUGAUGAAAGGGCAU) (10 nM, MSTUD0173, Sigma Aldrich), Slfn4 siRNA or the scrambled controls were transfected using Lipofectamine RNAiMAX transfection reagent (Thermo Fisher) for 48 hours. To induce Slfn4 expression, the cells were treated with 800 U/mL recombinant IFNα (R&D, #12 125–1) for 24 hours or at different time points.
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