Skip to main content
Fig. 5 | Journal of Neuroinflammation

Fig. 5

From: The impact of hyperpolarization-activated cyclic nucleotide-gated (HCN) and voltage-gated potassium KCNQ/Kv7 channels on primary microglia function

Fig. 5

Impact of functional HCN and KCNQ/Kv7 channels on the activation capacity of primary microglia. *p < 0.05; **p < 0.01; ***p < 0.001. Stimulation of microglia with LPS (10 ng/ml) with simultaneous blockade of the HCN or the KCNQ/Kv7 channel and resulting expression of pro-inflammatory markers. The HCN channel was either blocked pharmacologically with ZD7288 (30 μM) or by transfection of silencer® small interfering (si)RNA targeting HCN2-mRNA (siHCN2-RNA). XE-991 (30 μM) was used to block KCNQ/Kv7. a Inducible nitric oxide (NO) synthase (iNOS) expression was measured on RNA level by RT-qPCR (H(3) = 17.142, p = 0.001) and on b protein level by immunocytochemistry (H(3) = 19.626, p < 0.0001). c NO release was measured by Griess assay (μmol/l; H(3) = 23.102, p < 0.0001). Data were normalized to LPS-only stimulation. Characterization of the anti-inflammatory microglia phenotype by the expression of CD206, release of insulin-like growth factor 1 (IGF1), and measurement of the phagocytic activity, after treatment with IL4 (50 ng/ml) and simultaneous blockade of the HCN and KCNQ/Kv7 channel as described above. d Regulation of CD206 expression was measured on RNA level by RT-qPCR (H(3) = 12.755, p = 0.005). e Release of IGF1 was measured by ELISA (pg/ml; H(3) = 16.611, p = 0.001). f Zymosan engulfment indicated phagocytotic activity of microglia (H(2) = 7.501, p = 0.024). Data were normalized to IL4-only stimulation. g Representative images of the immunocytochemical staining for the microglia marker Iba1 (red), co-stained for iNOS (green), and Hoechst as a nuclear counterstain (blue); scale bars = 50 μm

Back to article page