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. 2005 Feb 21;201(4):647–657. doi: 10.1084/jem.20041611

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Copyright © 2005, The Rockefeller University Press

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Figure 7. — Knockdown and overexpression of TREM2 receptors in microglia by lentiviral vectors. (A) Gene transcript analysis of cultured microglia transduced with the shTREM2, shControl, wTREM2, or GFP2 vector. Gene transcripts for TREM2 and GAPDH were analyzed by RT-PCR. The lentiviral vector producing short hairpin RNAs specific for TREM2 induced complete knockdown of TREM2 gene transcripts. Gene transcripts for GAPDH and DAP12 served as an internal control and were not affected by the shTREM2 vector. Molecular weight marker: 100-bp ladder. (B) Protein expression of TREM2 was analyzed by a polyclonal antibody directed against TREM2 of cultured microglia transduced with the shTREM2 or shControl vector. Expression of TREM2 was not detectable by immunohistochemistry in microglia transduced with the shTREM2 vector. Bars, 20 μm. (C) Flow cytometry analysis of cultured microglial cells transduced either with a lentiviral vector producing short hairpin RNAs specific for TREM2 (shTREM2), irrelevant short hairpin RNAs (shControl), wTREM2, or control GFP (GFP2). shTREM2 transduction expression of TREM2 was almost undetectable, whereas wTREM2 transduction increased levels of TREM2 expression on microglia.