Review Article

M1 and M2 Functional Imprinting of Primary Microglia: Role of P2X7 Activation and miR-125b

Figure 2

Regulation of proinflammatory and anti-inflammatory mediators by P2X7 activation and miR-125b inhibition in SOD1-G93A microglia. SOD1-G93A primary microglia obtained from brain cortex of P0/P1 B6.Cg-Tg(SOD1-G93A)1Gur/J mice were prepared as previously described [15]. Then, 5 × 105 cells/well were plated and transfected with 20 nM scramble miRIDIAN hairpin inhibitor (ctr) or miR-125b miRIDIAN hairpin inhibitor (anti-125b) for 48 h and exposed to BzATP 100 μM for the indicated times. Expression levels of IL1β (a) (primer pairs: F 5′-GCAACTGTTCCTGAACTCAACT-3′; R 5′-ATCTTTTGGGGTCCGTCAACT-3′), IL4r (b) (F 5′-CGAGTTCTCTGAAAACCTC-3′; R 5′-CCATCTGGTATCTGTCTG-3′), Arg1 (c) (F 5′-CCACGGTCTGTGGGGAAAGCCAAT-3′; R 5′-CTGCCAGACTGTGGTCTCCACCCA-3′), and BDNF (d) (F 5′-CGGCGCCCATGAAAGAAGTA-3′; R 5′-AGACCTCTCGAACCTGCCCT-3′) were measured by qRT-PCR. Data are calculated using the method and GAPDH transcript as normalizer (F 5′-CATGGCCTTCCGTGTTCCTA-3′; R 5′-CCTGCTTCACCACCTTCTTGAT-3′). Bars represent mean ± sem of ≥3 independent experiments , , and versus untreated cells, by unpaired -test.
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