Journal: Journal of Neuroinflammation
Article Title: The impact of hyperpolarization-activated cyclic nucleotide-gated (HCN) and voltage-gated potassium KCNQ/Kv7 channels on primary microglia function
doi: 10.1186/s12974-020-01779-4
Figure Lengend Snippet: HCN channel and KCNQ/Kv7 channel expression on primary microglia. #p < 0.05, ##p < 0.01, ###p < 0.001 compared different experimental groups as marked by horizontal bar. a A representative image of primary microglia culture shows purity by staining of Iba1 (red). Hoechst stains all cell nuclei blue. Images were obtained with a fluorescence microscope, scale bars = 100 μm. b Expression of microglia marker Iba1 and P2RY12. Expression of microglial ion channels KCNK13, the HCN-subunits HCN1, HCN2, HCN3, and HCN4, as well as the KCNQ/Kv7 channel subunits KCNQ2, 3, and 5, on primary rat microglia in vitro measured by RT-qPCR. Microglia were untreated, activated with LPS (10 ng/ml) or IL4 (50 ng/ml; for Iba1: F(2, 6) = 23.571, p = 0.001, ω = 0.913; for P2YR12. F(2, 6) = 31.897, p = 0.001, ω = 0.934; for HCN2: F(2, 18) = 113.981, p < 0.0001, ω = 0.956; for HCN3: F(2, 8) = 22.573, p = 0.001, ω = 0.892; for KCNQ3: F(2, 11) = 11.674, p = 0.002, ω = 0.777; for KCNQ5: H (2) = 9.168, p = 0.01)
Article Snippet: Microglia were left unstimulated (control) or were stimulated with 10 ng/ml lipopolysaccharide (LPS derived from E. coli 0111: B4, Sigma Aldrich, St. Louis, USA) to activate microglia towards a pro-inflammatory phenotype, or alternatively stimulated with 50 ng/ml recombinant rat interleukin-4 (IL4; Peprotech, Hamburg, Germany) to polarize microglia to an anti-inflammatory phenotype [ ].
Techniques: Expressing, Staining, Fluorescence, Microscopy, Marker, In Vitro, Quantitative RT-PCR