Fig. 1

TrkA/B receptor expression in organotypic spinal cord cultures. a Low magnification immunofluorescence for SMI32 (green) of an organotypic spinal cord slice cultured for 2 weeks. The arrow indicates the ventral fissure, localizing the ventral horns. Note the co-cultured DRGs (arrow heads). Calibration bar 500 μm. b High magnification immunofluorescence labeling for microglia, astrocytes, and neurons, details of the ventral horn (left) or DRG (right) are shown. Left) green: Iba1 (microglia); red: GFAP (astrocytes). Right) green: GFAP (astrocytes); magenta: SMI-32 (neurons). Calibration bar 20 μm. c Western blot analysis of Tropomyosin receptor kinases A and B (TrkA and TrkB) expression in organotypic cultures stimulated with a cocktail of pro-inflammatory CKs (TNF-α, IL-1β and GM-CSF, 10 ng/ml each) for 4 or 6 h in the presence or in the absence of NGF-mimetic molecule MT2 (10 μM). GAPDH was used as house-keeping gene. Each well of the electrophoresis gel was loaded with an equal amount of proteins extracted from a pull of n = 5 organotypic slices. Note that TrkA and TrkB expression is not affected by treatments. This experiment is representative of 3 independent ones. d Immunofluorescence confocal and differential interference contrast (DIC) images for TrkA and TrkB expression in DRG neurons. Green: TrkA (left panel), TrkB (right panel). Red: SMI-32. Yellow: merge. Calibration bar 50 μm. e Immunofluorescence confocal images for TrkB and GFAP in the ventral horns. Green: TrkB; magenta: GFAP Calibration bar 20 μm