Figure 1

Targeting signal transducer and activator of transcription 3 (STAT3) in microglia by using STAT3^fl/fl;LysM-Cre^+/− mice. (a) Schematic representation of breeding strategy. The first offspring (F1) were obtained from a STAT3^fl/fl/LysM-Cre^+/+ crossing and STAT3^wt/fl;LysM-Cre^+/− (F1) were further mated with STAT3 floxed mice. The second offspring (F2) were divided into four groups: STAT3^wt/fl;LysM-Cre^−/−, STAT3^fl/fl;LysM-Cre^−/−, STAT3^wt/fl;LysM-Cre^+/−, and STAT3^fl/fl;LysM-Cre^+/−. (b) PCR analysis of STAT3 floxed (STAT3^fl/fl), wild-type (WT) (STAT3^wt/fl) and Cre genes in the second offspring. (c) Immunofluorescence staining for STAT3 (red), Iba-1 (green) and 4',6-diamidino-2-phenylindole (DAPI; blue) was performed in the prefrontal cortex (PFC) of the WT and knockout (KO) mice. Scale bar=40 μm. Scale bar of the enlarged image=10 μm. The graph shows intensity profiles illustrating STAT3/Iba-1 co-localization measurements. (d) Immunocytochemistry for STAT3 (green) and Iba-1 (red) was performed in the primary microglia cells of the WT and KO mice. Scale bar=50 μm. (e) The relative immunofluorescence intensity was used to represent protein levels of STAT3 and Iba-1 (STAT3; 1±0.058 vs 0.245±0.021, Iba-1; 1±0.075 vs 0.974±0.054 in primary microglia of the WT and KO mice, n=3 mice/group, respectively). (f) Western blot analysis of STAT3, Iba-1, PSD95, and GFAP in primary microglia, neurons, and astrocytes of the WT and KO mice. Data are means±SEM, and *p<0.05, **p<0.01, and ***p<0.001. Representative data from three independent experiments are shown.

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