Raw lucia

Manufactured by InvivoGen

Sourced in United States

The RAW-Lucia- is a cell line derived from the human monocytic cell line THP-1. It was engineered to stably express the Lucia luciferase reporter gene, which can be used as a readout for monitoring cellular activation and signaling pathways.

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Lab products found in correlation

Raw lucia isg mda5 ko, by InvivoGen (2 mentions) Raw lucia isg rigi ko, by InvivoGen (2 mentions) Quanti luc, by InvivoGen (2 mentions) Thp1 dual cells, by InvivoGen (1 mentions) Thp1 dual ko sting cells, by InvivoGen (1 mentions) Phospho irf3 ser396, by Cell Signaling Technology (1 mentions) Diabzi compound 3, by Selleck Chemicals (1 mentions) Quanti luc solution, by InvivoGen (1 mentions) Quanti blue solution, by InvivoGen (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Skh 1e, by Charles River Laboratories (1 mentions) Normocin, by InvivoGen (1 mentions) Raw lucia isg cell line, by InvivoGen (1 mentions) Zeocin, by InvivoGen (1 mentions) C57bl 6, by Charles River Laboratories (1 mentions) Female balb c mice, by Charles River Laboratories (1 mentions)

4 protocols using raw lucia

STING Pathway Modulation Assays

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THP1-Dual cells (Cat. code: thpd-nfis), THP1-Dual KO-STING cells (Cat. code: thpd-kostg), 293T-Dual hSTING-H232 cells (Cat. code: 293d-h232), 293T-Dual hSTING-R232 cells (Cat. code: 293d-r232), Raw-Lucia (Cat. code: rawl-isg) and Raw-Lucia-KO-STING (Cat. code: rawl-kostg) were purchased from InvivoGen (San Diego, USA). All the cells were incubated following manufacturer’s protocols at 37 ^oC in a humidified atmosphere of 5% CO₂. QUANTI-Luc solution (Cat. code: rep-qlc2) and QUANTI-Blue solution (Cat. code: rep-qbs2) were purchased from InvivoGen (San Diego, USA). Antibodies against phospho-TBK1 (Ser172) (Cat. code: 5483S), TBK1 (Cat. code: 3504S), phospho-IRF3 (Ser396) (Cat. code: 4947S), IRF3 (Cat. code: 4302S), and GAPDH (Cat. code: 5174S) were purchased from Cell Signaling Technology (Beverly, MA). diABZI-compound 3 (Cat. code: S8796) and MSA-2 (Cat. code: S9681) were purchased from Selleck (Houston, USA).

Xie Z., Wang Z., Fan F., Zhou J., Hu Z., Wang Q., Wang X., Zeng Q., Zhang Y., Qiu J., Zhou X., Xu H., Bai H., Zhan Z., Ding J., Zhang H., Duan W., Yu X, & Geng M. (2022). Structural insights into a shared mechanism of human STING activation by a potent agonist and an autoimmune disease-associated mutation. Cell Discovery, 8, 133.

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Quantifying IRF Activation in M. tuberculosis Infection

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RAW-Lucia-ISG, RAW-Lucia-ISG-MDA5-KO, and RAW-Lucia-ISG-RIGI-KO (InvivoGen) cells were generated from the murine RAW 264.7 macrophage cell line by stable integration of a secreted Lucia luciferase reporter under the control of an ISG54 minimal promoter and 5 IFN-stimulated response elements, thus allowing IRF activation to be monitored by levels of Lucia luciferase in the cell culture supernatant. In brief, the cells were inoculated with the M. tuberculosis H37Rv strain at an MOI of 5 or 20 for 2 hours and then washed with DMEM and further cultured in complete DMEM. After 24 hours of incubation, supernatants were collected and assayed for the level of IRF induction by determination of the activity of Lucia luciferase using QUANTI-Luc (InvivoGen) according to the manufacturer’s protocol.

Bullen C.K., Singh A.K., Krug S., Lun S., Thakur P., Srikrishna G, & Bishai W.R. (2023). MDA5 RNA-sensing pathway activation by Mycobacterium tuberculosis promotes innate immune subversion and pathogen survival. JCI Insight, 8(20), e166242.

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Murine Cancer Cell Culture and Maintenance

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All animal procedures were approved by Massachusetts Institute of Technology Committee on Animal Care. Six- to eight-week-old SKH1-E, C57BL/6, and BALB/c female mice were purchased from Charles River Laboratories Inc. The mouse breast cancer cell line 4T1 and melanoma cell line B16F10 were purchased from American Type Culture Collection. The RAW-Lucia ISG cell line was purchased from InvivoGen Inc. KPC (LSL-Kras^G12D/+; LSL-Trp53^R172H/+; Pdx-1-Cre) pancreatic cancer cells were kindly given by Dr. Serguei Kozlov (Frederick National Laboratory of Cancer Research). Cells were cultured in Dulbecco’s modified Eagle medium (DMEM, B16F10), DMEM/F12 (KPC), and RPMI 1640 (4T1) supplemented with 10% fetal bovine serum (FBS), penicillin (100 units/mL), and streptomycin (100 μg/mL) at 37 °C and with 5% CO₂. Cell culture media and antibiotics were purchased from Invitrogen. RAW-Lucia ISG cells were cultured in DMEM supplemented with 2 mM L-glutamine, 10% FBS, 100 μg/ml Normocin (InvivoGen), and 200 μg/ml Zeocin (InvivoGen).

Lu X., Miao L., Gao W., Chen Z., McHugh K.J., Sun Y., Tochka Z., Tomasic S., Sadtler K., Hyacinthe A., Huang Y., Graf T., Hu Q., Sarmadi M., Langer R., Anderson D.G, & Jaklenec A. (2020). Engineered PLGA microparticles for long-term, pulsatile release of STING agonist for cancer immunotherapy. Science translational medicine, 12(556), eaaz6606.

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Quantifying IRF Activation in M. tuberculosis Infection

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