Additional file 1: Figures S1–S6. Figure S1. of Sildenafil, a cyclic GMP phosphodiesterase inhibitor, induces microglial modulation after focal ischemia in the neonatal mouse brain

Outline of the experimental procedure in P9 C57Bl/6 mice subjected to pMCAo. Figure S2. A: mean blood flow velocities (BFVs) in the left and right ICA (lICA, rICA) and basilar trunk (BT) in basal conditions and 15 min after pMCAo. *, #p < 0.05 vs basal for each artery. B Variation in arterial blood flow (BF) in P9 mice subjected to pMCAo and administered with PBS (white bars) and/or sildenafil (15 mg/kg, black bars). Figure S3. cGMP dosage in the ipsilateral hemisphere in the control (PBS-treated naive mice, n = 5) and in sildenafil- (10 mg/kg) treated mice, at 1 and 3 h (h) after treatment. ***p < 0.001 vs PBS. Figure S4. Quantification of GFAP-positive cells in the penumbra at 72 h and 8 days after pMCAo in PBS- and sildenafil-treated animals (n = 8 per group). Figure S5. Immunohistochemistry for astrocyte (GFAP; A, B) and macrophage (tomato lectin (TL) C–D)/microglia (Iba-1; E, F) in PBS-treated animals (A, C, and E) and sildenafil 10 mg/kg (B, D, and F) at 72 h (A–D) and 8 days (E, F) after pMCAo in mouse pups. Figure S6. 3D confocal reconstruction of a microvessel stained with the 54 kDa Glut-1 protein (green) and microglia/macrophage stained with MRC-1 protein (M2-like marker). (PDF 1359 kb)