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cmi1838-sup-0001-fS1.tiff298K

Fig. S1. Inhibition of galectin-3 binding to N. meningitidis.

A. Flow cytometric quantification of galectin-3 (10 μM) binding to N. meningitidis strain MC58 (2 × 107 bacteria) in presence of lactose or sucrose. Lactose at a concentration of 300 mM partially inhibits galectin-3 binding (**P <0.01, P = 0.0021), while 300 mM sucrose does not affect binding to fixed bacteria.

B. Quantification of galectin-3 binding to fixed bacteria in presence of 300 mM lactose and increasing concentrations of galectin-3 (0.033, 0.333, 1.6, 3.3, 6.6, 10 μM). Galectin-3 binding is inhibited by 300 mM lactose up to a galectin-3 concentration of 10 μM.

cmi1838-sup-0002-fS2.tiff434K

Fig. S2. Association of meningococci to PMA-differentiated THP-1 macrophages analysed by bacterial recovery following infection for 6 h with bacteria pre-incubated with 3.3 μM galectin-3 (black bars) or PBS (grey bars).

A. The association of bacteria pre-incubated with galectin-3 is significantly increased compared to bacteria pre-incubated with PBS (*P = 0.0140). Data are from three different experiments performed in duplicate.

B. Uptake of meningococci by THP-1 macrophages shown as the percentage relative to the total number of bacteria in the inoculum. There is no significant difference in uptake of bacteria following pre-incubation of bacteria with galectin-3 or PBS. Data are the mean of three independent experiments performed in duplicate and error bars show the standard deviation. Statistical significance was calculated using a paired, one-tailed t-test (GraphPad Prism).

cmi1838-sup-0003-fS3.tiff421K

Fig. S3. Internalization of bacteria by THP-1 macrophages. Uptake of meningococci by THP-1 macrophages (shown as a percentage of the inoculum) 1 h post inoculation and following gentamicin treatment to kill extracellular bacteria. The percentage of bacteria which are internalized is not significantly different following pre-incubation with PBS or galectin-3 (one tailed, paired Student's test; P = 0.1031). Data are the mean of six different experiments and error bars indicate the standard deviation.

cmi1838-sup-0004-fS4.tiff2K

Fig. S4. Association of wild-type MC58 and MC58ΔlgtB to primary monocytes. The actual percentage of bacterial association is measured in relation to the initial bacterial inputs. The MC58ΔlgtB strain adheres to monocytes at similar levels to the wild-type strain MC58. Data are the mean of nine different experiments and error bars show standard deviation.

cmi1838-sup-0005-fS5.tiff511K

Fig. S5. Flow cytometric quantification of human and mouse galectin-3 binding to N. meningitidis strain MC58. Fixed bacteria were incubated with increasing concentrations of galectin-3 (0.033, 0.333, 1.6, and 3.3 μM). Galectin-3 from each species binds to N. meningitidis. Mean and standard deviation is shown and data is from one representative experiment.

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