Fig. 3

Hippocampal and cortical microglia cell reactivity, age-related effects and dynamic activation in chronic hypertensive states. (a, e) Identification of microglia derived from the hippocampus and cortex of SHRSP rats via FACS analysis. Single live cells were identified by the exclusion of doublets, cell debris and gating of viable cells (Additional file 4: Fig. 3). Main populations of cells were identified through Forward Scatter light (FSC) and CD45. Leukocytes were selected based on their size and CD45 expression. Higher FSC CD45⁺ events were further gated as CD45⁺ and CD11b/c⁺ positive events and further classified by their expression of P2Y12R and CD11b/c. Double positive events were classified as microglia in early chronic hypertension and (b, f) late chronic hypertension. (c) Frequency and count of microglia cells in the hippocampus and (g) the cortex, median fluorescence intensity (MFI) for FSC was used to investigate microglia size. (d, h) Bar charts showing the fold change in MFI in each surface antigen investigated in hippocampal and cortical microglia. Individual values shown in the graph represent mean ± SEM. p-values: * ≤ 0.05; ** for p ≤ 0.01; *** for p ≤ 0.001; **** for p ≤ 0.0001