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. Author manuscript; available in PMC: 2014 Jun 23.
Published in final edited form as: Stem Cells. 2008 Jun 26;26(10):2735–2745. doi: 10.1634/stemcells.2008-0212

Figure 6.

Figure 6

Targeted intracellular peptide and gene delivery with SPARC-mimicking peptides. (A): Dose-response effect of mPep-GG-D(KLAKLAK)2 on mouse 3T3–L1 preadipocytes induced toward adipocyte differentiation. Cells were treated with increasing concentrations of mPep-GG-D(KLAKLAK)2 or negative controls (D(KLAKLAK)2 untargeted or fused with an unrelated peptide) for 24 hours. Apoptosis was assessed by the WST-1 assay. Error bars: SEM from triplicate wells. (B, C): Micrographs of 3T3–L1 cells in (A) after 24 hours of treatment: 0.1 mM mPep-GG-D(KLAKLAK)2 induced apoptosis (B), whereas untargeted D(KLAKLAK)2 had no effect on 3T3–L1 differentiation (C). (D–G): Delivery of an antilipogenic transgene (Ucp1 expressed from AAVP) with SPARC-mimicking peptides into 3T3–L1 cells induced toward adipocyte differentiation. AAVP-Ucp1 displaying mPep efficiently transduced preadipocytes, which resulted in Ucp-1 expression detected (arrow) with anti-Ucp1 antibody (red immunofluorescence) 5 days post-transduction (D), whereas untargeted AAVP-Ucp1 did not transduce preadipocytes (E). Micrographs taken 7 days post-transduction demonstrate that expression of Ucp-1 upon mPep-targeted AAVP-Ucp1 delivery prevented adipocyte differentiation of 3T3–L1 cells (F), whereas the untargeted AAVP-Ucp1 vector did not prevent their differentiation into adipocytes (G). Scale bar % 5 µm. Abbreviations: AAVP, adeno-associated virus phage; OD, optical density.