Fig. 5

CD73 deficiency promoted microglial M1 polarization and inhibited microglial M2 polarization in BV2 cells, while CD73 overexpression had an opposite effect. After small interference RNA (siRNA) or pcDNA3.1 transfection for 24 h, BV2 cells were challenged with LPS or IL-4 for another 8 h to activate M1 or M2 activation. Then, Western blot and real-time PCR were applied to determine the expression of M1 or M2 markers in BV2 cells under different culture conditions. a Real-time PCR estimates the efficiency of CD73 interference and overexpression in BV2 cells 24 h after transfection. b, c Representative immunoblotting of CD73 expression and quantification 24 h after transfection. d, e, h, i After administration of LPS or IL-4, microglial M1 polarization markers (TNF-α, IL-1β, IL-6, iNOS, CD86) or M2 polarization markers (Arg1, IL-10, CD206) mRNA expression were identified by real-time PCR in CD73 downregulated or upregulated BV2 cells. f, g, j, k Representative Western blotting and quantification of iNOS or Arg1 expression induced by LPS/IL-4 in CD73 different expression BV2 cells. *p < 0.05, **p < 0.01, ***p < 0.001. Data are shown as the mean ± SD from four independent experiments