FIGURE 10:

Effects of AUF1-RGG peptides on endogenous VEGF protein levels. Cultured RAW-264.7 cells were transfected with the empty parental vector, pcDNA-3.1 (PC), the AUF1-QRGG peptide vector (QRGG), or the AUF1-6+8 peptide vector (6+8). After 4 h, cells were treated with LPS for 18 h. Lysates were probed for VEGF and α-tubulin. (A) Immunoblot for VEGF or α-tubulin. (B) Levels of VEGF protein normalized to α-tubulin for six experiments. As compared with pcDNA-3.1, both AUF1-QRGG (p < 0.002) and AUF1-6+8 (p < 0.003) significantly decreased VEGF protein. (C) Effects of AUF1 peptides on VEGF-3′ UTR reporter activity. Cells (RAW-264.7) were transfected with VEGF-3′ UTR-Luc and were cotransfected with pcDNA-3.1, AUF1 (AUF1-p37), or AUF1-RGG peptides (Ex6+8 or QRGG). The luciferase activity in cell lysates from triplicate wells was read after 24 h. p < 0.0001 for pcDNA vs. AUF, AUF1-Ex6+8, or AUF1-QRGG. No other comparators were significantly different. Results are normalized to pcDNA treatment and are representative of two or more experiments.