Extended Data Fig. 2: LRRK2 is not involved in NOD2 signalling in MODE-K cells.
From: LRRK2 is not required for lysozyme expression in Paneth cells
a) MODE-K cells treated with TLR2 and NOD2 agonists Pam3CSK4 (1 µg/ml), MDP (10 µg/mL) with or without LRRK2 inhibitor MLi-2 (100 nM) for 24 h. Immunoblot of pSer935LRRK2, total LRRK2, pThr73 Rab10, total Rab10 and loading control α-tubulin from one experiment. b) Equal numbers of MODE-K cells were treated for 24 h with or without MLi-2 (100 nM), and with or without TLR2 and NOD2 agonists Pam3CSK4 (1 µg/ml), MDP (10 µg/mL). Supernatants were collected and used to measure IL-6 by ELISA from 3 independent experiments. Significance was determined by two-way ANOVA and Å Ãdák’s multiple comparisons test. c) Signalling downstream of NOD2/TLR2 was investigated in MODE-K cells treated for 4hrs with or without MLi-2 (100 nM), and with or without TLR2 and NOD2 agonists Pam3CSK4 (1 µg/ml), MDP (10 µg/mL). Immunoblotting for pSer536 p65, total p65, pThr180/Tyr182 p38, total p38 and tubulin as a loading control are shown. d) Quantification of p-p38 normalised to the loading control from 4 independent replicates is shown. Significance was determined by two-way ANOVA and Å Ãdák’s multiple comparisons test. Mean +/- s.d. and individual replicates are represented as bars, error bars and dots, respectively.
