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. 2011 Nov 18;9(1):51–58. doi: 10.7150/ijms.9.51

Figure 4.

Figure 4

DLS inhibits chemokine- mediated signaling in CCR5-transfected 3T3 cell lines. Approximately 200,000 3T3.T4.CCR5 cells were grown overnight per well in 6-well plate. On the following day, the media was replaced with 2ml of serum free medium. After 2-3 hrs, DLS (20μM) or the CCR5 specific antagonist, TAK779 (2μM), were added to respective wells, incubated for 30 minutes at 37°C after which 10nM of RANTES (A) or MIP-1β (B) was added to indicated wells. The cells harvested in RIPA after washing with cold PBS at the indicated times. 30μg of protein run in 4-20% Bis-Tris gel, transferred and probed with mouse anti-pERK antibody (upper panel) stripped and again probed with rabbit anti-total ERK antibody (lower panel). The results reveal that DLS partially blocks RANTES induced ERK activation but surprisingly does not block MIP-1β-induced ERK activation in 3T3.T4.CCR5 cells.