Research Paper Volume 11, Issue 3 pp 908—920

Genetical modification on adipose-derived stem cells facilitates facial nerve regeneration

Figure 3. Preparation of ASCs-miR-449 and ASCs-shPLOD1. (A) Schematic of AAVs. 1. an AAV carrying short-hairpin small interfering RNA for PLOD1 (shPLOD1) and a GFP reporter under control of a CMV promoter. 2. an AAV carrying miR-449 and a GFP reporter under control of a CMV promoter. 3. an AAV carrying GFP and scr. (B) Transduced rat ASCs were purified based on GFP expression by flow cytometry. (C) RT-qPCR for PLOD1 levels in ASCs-miR-449, ASCs-shPLOD1 and control ASCs-scr. (D) Western blotting for PLOD1 levels in ASCs-miR-449, ASCs-shPLOD1 and control ASCs-scr. (E) Oil red O staining to evaluate adipogenic induction. (F) Alcian blue staining to evaluate chondrogenetic induction. (G) Von Kossa staining to evaluate osteogenic induction. *p<0.05. NS: non-significant. N=5. Scale bars are 50µm.