Fig. 2

Reactive microglia in the ACC are required for nicotine-induced allodynia. A Schematic timeline of experiment. B Representative imaging of Iba-1-labelled microglia after intra-ACC ACSF in vehicle mice and intra-ACC ACSF or MINO in nicotine-treated mice. Scale bars, 40 μm (overview) and 10 μm (Zoom and Rendering). C Quantification of Iba-1⁺ number, intensity, soma size and Imaris-based semi-automatic quantification of Iba-1⁺ microglia morphometry in the ACC. D Changes of PWT and PWL in vehicle- and nicotine-treated mice with intra-ACC ACSF or MINO. E Schematic timeline of experiment. F Representative imaging of Iba-1-labelled microglia after intra-ACC ACSF in vehicle mice and intra-ACC ACSF or Lip in nicotine-treated mice. Scale bars, 40 μm (overview). G (Left) Quantification of Iba-1⁺ number in the ACC. (Middle and right) Changes of PWT and PWL in vehicle- and nicotine-treated mice with intra-ACC ACSF or Lip. Data were presented as mean ± SEM or Median (IQR). *p < 0.05; **p < 0.01; ***p < 0.001; ns not significant. Details of the statistical analyses are presented in Additional file 1: Table S1