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. 2020 May 1;11:2148. doi: 10.1038/s41467-020-15978-3

Fig. 5. AGE products stimulate MD2-dependent inflammatory responses.

Fig. 5

a AGE product formation in MPMs exposed to HG in the presence or absence of serum. MPMs were exposed to 33 mM glucose for different time periods in media containing 0 or 10% FBS. Levels of AGE products were determined in conditioned medium by ELISA [means ± SEM; n = 4 examinations]. b AGE product formation in H9C2 cells exposed to HG in the presence or absence of serum. H9C2 cells were exposed to 33 mM glucose for different time periods in media containing 0 or 10% FBS. Levels of AGE products were determined in conditioned medium by ELISA [means ± SEM; n = 4 examinations]. c Representative immunoblot showing co-immunoprecipitation of MD2-TLR4 complex in MPMs exposed to 33 μg/mL AGE-BSA [n = 6]. d Representative immunoblot showing co-immunoprecipitation of MD2-TLR4 complex in H9C2 cells exposed to 33 μg/mL AGE-BSA [n = 3]. e Levels of TNF-α and IL-6 in condition media of MPMs exposed to 33 μg/mL AGE-BSA for 24 h. MPMs isolated from WT or MD2KO mice were tested [means ± SEM; n = 4 examinations]. f Levels of Tnfa and Il6 mRNA in H9C2 cells exposed to 33 μg/mL AGE-BSA for 6 h. H9C2 cells were transfected with control siRNA or siRNA targeting MD2 (siMD2) before treatments [means ± SEM; n = 6 examinations]. Source data are provided as a Source Data file. P-values by one-way ANOVA in a, b, e, f followed by Tukey’s post hoc test are indicated.