Research Paper Volume 12, Issue 8 pp 6558—6569

Figure 1. Preparation of AAVs that deplete DNMT1 and TGFβ1. (A) Schematic to show the structure of AAVs carrying shRNA for DNMT1 (shDNMT1) or shRNA for TGFβ1 (shTGFβ1) under a macrophage-specific CD68 promoter. The control AAV carried a scramble sequence under the CD68 promoter. The constructs were connected to a GFP reporter by p2A to allow co-expression by one promoter. (B, C) DNMT1 (B) and TGFβ1 (C) levels were examined in 3 mouse macrophage lines, Raw264.7, J774A.1 and IC-21, by RT-qPCR. (D–G) IC-21 was transduced with different AAVs. DNMT1 levels were determined by RT-qPCR (D) and by Western blot (E). TGFβ1 levels were determined by RT-qPCR (F) and by ELISA (G). *p<0.05. NS: non-significant. N=5.