Fig. 4
From: Microglia amplify inflammatory activation of astrocytes in manganese neurotoxicity
Microglia uptake 70% of Mn present in media. a Schematic diagram outlining the procedure for microglia-conditioned media (MCM) experiments. The amount of Mn remaining in MCM media was assessed via microglia cell uptake via cellular fura-2 manganese extraction assay (CFMEA) (b, c) or by measuring Mn levels in media (d). b Cell-free Mn-fura-2 saturation binding standard curve generated to calculate Mn uptake in microglia over a 24-h period. Data is represented as mean levels ± SD at %MAX (x-axis) and log10 scale of MnCl2 concentration (y-axis) with power curve (red line) used for %MAX less than 50% and logarithmic curve (blue line) used for %MAX greater than 50%. c The calculated amount of Mn uptake (y-axis) per MCM experiment measured via CFMEA in cultured microglia cells exposed to increasing doses of MnCl2 (0–100 μM; x-axis) over 24 h. Data is represented as mean Mn uptake per 2.5 × 105 cells ± SEM (one-way ANOVA; asterisk indicates significance from control; *p < 0.05). d The levels of Mn in MCM were measured via ICP-MS at time 0 and 24 h post treatment. Data are presented as the mean micromolar concentration of Mn ± SEM (one-way ANOVA; asterisks indicate significance from control; ****p < 0.0001). e Schematic diagraming procedure for microglia-astrocyte co-culture experiments