Overexpression and knockdown of MFG-E8 in microglial cells after transfection with a lentivirus.

Four different transfections were used: M+, MFG-E8 overexpression using pGC-FU-MFG-E8; M-, MFG-E8 negative control for overexpression using the pGC-FU vector; R+, Knockdown expression using pFU-GW-RNAi-MFG-E8; R- negative control for knockdown expression using the pFU-GW Vector. (A) The transfection rate was over 80% in all experiments. (B) qRT-PCR analysis indicated that pGC-FU-MFG-E8 significantly enhanced microglial MFG-E8 expression, and pFU-GW-RNAi-MFG-E8 effectively knocked it down. The negative controls had no effect. (C) Western blot analysis of the cell lysate from the transfected microglial cells confirmed that MFG-E8 expression (about 47 kDa) was significantly enhanced or knocked down. Scale bar: 50 µm. ** P<0.01.