Investigating LRRK2 Kinase Inhibition in Microglia

Zymosan A from Saccharomyces cerevisiae (Sigma, St. Louis, MO, USA, #Z4250) and MLi-2 from Merck (Tocris, Bristol, UK, #5756) were used for these experiments as TLR2 inflammatory stimuli and an LRRK2 kinase inhibitor, respectively. For immunoblots, primary microglia from wild-type and G2019S-LRRK2 Tg mice were treated with 200 µg/mL zymosan and 1 µM MLi-2 for 4 h and 24 h, and for RNA-Seq, they were treated with the same concentration of zymosan and MLi-2, but for 24 h only. Zymosan dose was chosen based on previous studies that have used 200 μg/mL concentration of zymosan in immune cells, such as in BMDMs [16 (link),20 (link)]. For MLi-2, previous studies have used a 1 mM concentration of MLi-2 in vivo [21 (link)] and a 0.5 mM concentration of MLi-2 in vitro [22 (link)].