Figure legends
Figure 1 Roles of RAGE in modulating tick-bite induced inflammation at different phases of tick infestations. (A) Histological feature of the tick-bitten areas or normal (control) skin. Ticks were allowed to feed on C57BL/6 or RAGE -/- mice, and the same mice were used for repeated tick infestations. Mice were euthanized after repletion of tick in each infestation (n=3 for each feeding phase). Skin from the attachment site of ticks was collected and preserved in 4% paraformaldehyde added with 0.1% gluteraldehyde overnight at 4 °C under gentle shacking. Thin (5 μm) sections were prepared and subjected to H&E. (B) Comparative features of cellular infiltration in tick-bitten skins. Ticks were allowed to feed on C57BL/6 or RAGE -/- mice in the same manner and subjected to histopathological analysis. Inflammatory cells per focus were counted by two independent blind investigators. **, P<0.001; NS, non-significant.
Figure 2 Upregulation of RAGE in tick bite areas. Ticks were allowed to feed on C57BL/6 or RAGE -/- mice, and the same mice were used for repeated tick infestations. Mice were euthanized after repletion of tick in the tertiary infestation (n=3). Skin from the attachment site of ticks was collected and preserved in 4% paraformaldehyde added with 0.1% gluteraldehyde overnight at 4 °C under gentle shacking. Thin (5 μm) sections were prepared and subjected to immunofluorescence using anti-RAGE as mentioned in the methodology section.
Figure 3 Detection of RAGE ligands at the tick bite areas.Ticks were allowed to feed on C57BL/6 orRAGE -/- mice, and the same mice were used for repeated tick infestations. Mice were euthanized after repletion of tick in the tertiary infestation (n=3). Skin from the attachment site of ticks was collected and preserved in 4% paraformaldehyde added with 0.1% gluteraldehyde overnight at 4 °C under gentle shacking. Thin (5 μm) sections were prepared and subjected to immunohistochemistry using anti-S100A/8 or anti-S100B as mentioned in the methodology section.
Figure 4 RAGE dictates ILC2s during ticks’ feeding. Ticks were allowed to feed on C57BL/6 or RAGE -/- mice, and the same mice were used for repeated tick infestations. Mice were euthanized after repletion of tick in the tertiary infestation (n=3). Skin from the attachment site of ticks was collected and preserved in 4% paraformaldehyde added with 0.1% gluteraldehyde overnight at 4 °C under gentle shacking. Thin (5 μm) sections were prepared and subjected to immunohistochemistry using anti-CD44 or anti-MPO as mentioned in the methodology section.
Figure 5 Tick-bite induced eosinophilia is governed by RAGE.Ticks were allowed to feed on C57BL/6 orRAGE -/- mice, and the same mice were used for repeated tick infestations. Blood were collected different feeding phase at different time points (day 0-day 5) from tail tip and examined.