Transcriptomic changes in RAW264.7cells exposed to bacteriallipopolysaccharide (LPS) and/or thenitrone compound 5,5-dimethyl-1-pyrroline N-oxide (DMPO)DMPO

DARIO C RAMIREZ; GOMEZ MEJIBA, SANDRA;

MENDELEY DATA

Elsevier BV

Año: 2025

2692-8205

The main hypothesis to be tested is that DMPO causes transcriptomic changes in macrophages (RAW264.7 cell line) to make them resistant to activation with bacterial lipopolysaccharide (LPS) , chromatographically purified from E. coli 055:B5. The main objective was to characterize the overall trasncriptomic changes caused in RAW264.7 cells exposed to DMPO and/or LPS. RAW264.7 cells (ATCC TIB-71, passage 3-4) were cultured in T-25 flask in DMEM culture medium with high glucose and 5% heat inactivated fetal calf serum (GIBCO). After reaching a 70% confluence, monolayers were rinsed and incubated in medium alone, medium+DMPO (50mM), medium+LPS (1ng/ml), medium+DMPO+LPS. After incubation for 6h, in 5% CO2 at 37oC, under the 4 different conditions the medium was removed, the mnolayers rinsed with PBS. Following the cell monolayers were harvested in 1ml TRIZOL to purify total RNA. RNA integrity was assessed by capilar electrophoresis (Eukaryote Total RNA Nano). This experiment