Rgs2 regulates MCP-1 expression in MDSCs.

<p>MDSCs were isolated from 3LL tumor tissues of wild type or Rgs2−/− mice by magnetic sorting after digestion of the tissues with hyaluronidase and collagenase. (A) Protein lysates from the isolated cells were analyzed using a cytokine array. Each cytokine is detected in duplicate, and intensity was determined using ImageJ software. Positive controls provided on the array were used for normalization. This experiment was performed twice. (B) RNA was extracted from the isolated MDSCs, and analyzed by real time PCR. Beta-actin was used as an internal control. This experiment was repeated 3 times. (C) MDSCs were isolated from tumors of wild type and Rgs2 null mice, and cultured for 48 hours. Culture medium was assayed for MCP-1 protein by ELISA. This experiment was performed in duplicate and repeated 3 times. * p<0.05, ** p<0.00005. (D) MDSCs were isolated from normal spleen and 3LL tumor tissues of wild type mice. Rgs2 and MCP-1 levels were measured by real time PCR. Beta-actin was used as an internal control. This experiment was repeated 2 times. * p<0.05, ** p<0.005.