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Additional Supporting Information may be found in the online version of this article.

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GLIA_21423_sm_SuppFig1.TIF3698KSupporting Information Figure 1. TLR2 loss leads to elevated IL-12 family cytokine production in macrophages upon exposure to intact S. aureus. Thioglycollate-elicited peritoneal macrophages from TLR2 WT and KO mice were seeded at 2 × 105 cells per well in 96-well plates and incubated overnight. After 24 h, cells were exposed to 107 cfu heat-inactivated S. aureus for 24 h, whereupon IL-12p40 (A), IL-12p70 (B), IL-27 (C) and CCL2 (D) production was quantitated by ELISA. Significant differences between TLR2 KO versus WT macrophages are indicated by asterisks (*, p < 0.001). Results are reported as the mean ± SD of three independent wells for each experimental treatment and were identical across three separate experiments.
GLIA_21423_sm_SuppFig2.TIF3130KSupporting Information 2. Exaggerated IL-12p40 production by TLR2 KO macrophages is conserved upon exposure to various streptococcal and staphylococcal species. Thioglycollate-elicited peritoneal macrophages from TLR2 WT and KO mice were seeded at 2 × 105 cells per well in 96-well plates and incubated overnight. After 24 h, cells were exposed to 2 × 105 cfu/well of live Gram-positive bacteria for 6 h, whereupon IL-12p40 production was quantitated by ELISA. Significant differences between TLR2 KO versus WT macrophages are indicated by asterisks (*, p < 0.05). Results are reported as the mean ± SD of two independent wells for each experimental treatment and were identical across two separate experiments.
GLIA_21423_sm_SuppFig3.TIF977KSupporting Information 3. Gram-negative bacteria do not augment IL-12p40 production in TLR2 KO macrophages. Thioglycollate-elicited peritoneal macrophages from TLR2 WT and KO mice were seeded at 2 × 105 cells per well in 96-well plates and incubated overnight. After 24 h, cells were exposed to 2 × 105 cfu/well of live Gram-negative bacteria for 6 h, whereupon IL-12p40 production was quantitated by ELISA. Results are reported as the mean ± SD of three independent wells for each experimental treatment and were identical across two separate experiments.
GLIA_21423_sm_SuppFig4.TIF786KSupporting Information 4. TLR9 blockade prevents exaggerated IL-12p40 expression in TLR2 KO macrophages. Thioglycollate-elicited peritoneal macrophages from TLR2 WT and KO mice were seeded at 2 × 105 cells per well in 96-well plates and incubated overnight. After 24 h, cells were exposed to heat-inactivated S. aureus (107 cfu/well) ± the inhibitory CpG ODN 2088 for 24 h, whereupon IL-12p40 production was quantitated by ELISA. Significant differences between TLR2 KO versus WT microglia are indicated by asterisks (*, p < 0.05), whereas differences between TLR2 KO microglia treated with S. aureus only versus S. aureus ± ODN 2088 are indicated by hatched signs (#, p < 0.05). Results are reported as the mean ± SD of three independent wells for each experimental treatment and were identical across two separate experiments.

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