Gene expression
BMDM, THP-1 cells, or human monocytes were cultured with 200 ng/ml LPS
for 4 hours and then treated with 10 or 100 μM LMT503 for 18 hours at
37°C. To investigate gene expression in colon macrophages, mononuclear
cells were isolated from whole colon tissue as described previously (Kim
et al., 2018). Macrophages were further purified with MagniSort Mouse
F4/80 Positive Selection Kit (Invitrogen, Carlsbad, CA, USA). Whole RNAs
were extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA),
followed by cDNA synthesis using a SuperScript II Reverse Transcriptase
(Invitrogen, Carlsbad, CA, USA). Gene expression was measured by
StepOnePlus RT-PCR (Applied Biosystems, Waltham, MA, UNA). Primers used
for PCR are listed in Supplementary Table 1. Each sample was run in
triplicate and three independent experiments were performed for each
sample.