Figure 4.

The plots of the real time detection and amplification of mRNA of CXC chemokine receptor 4 (CXCR4) in unstimulated, and in interleukin (IL)-4- and IL-10-stimulated, CD4⁺ T lymphocytes at different time intervals. (a) Green plots represent the amplification of mRNA of CXCR4 in unstimulated CD4⁺ T lymphocytes; blue, orange and red plots represent the amplification of mRNA of IL-4-stimulated CD4⁺ T lymphocytes at 4 hr, 16 hr and 24 hr, respectively; black plots represent the amplification of standard DNA template (2·0 × 10⁴ copies) with a housekeeping gene (β-actin). C_T values (representing the polymerase chain reaction cycle at which an increase in reporter fluorescence signal above baseline was first detected) were 18·1 for standard DNA template; 24·7 for CXCR4 mRNA in unstimulated CD4⁺ T lymphocytes; 22·8 for CXCR4 mRNA in IL-4-stimulated CD4⁺ T lymphocytes at 4 hr; 18·6 for 16 hr; and 22·8 for 24 hr, respectively. (b) Green plots represent the amplification of CXCR4 mRNA in unstimulated CD4⁺ T lymphocytes; blue, orange and red plots represent the amplification of mRNA of IL-10-stimulated CD4⁺ T lymphocytes at 4 hr, 16 hr and 24 hr, respectively; black plots represent the amplification of standard DNA template (2·0 × 10⁴ copies) with a housekeeping gene (β-actin). C_T values were 24·2 for CXCR4 mRNA in IL-10-stimulated CD4⁺ T lymphocytes at 4 hr; 34·9 for 16 hr; and 24·7 for 24 hr, respectively. (c) Demonstration of the linear relationship between C_T and log starting quantity (S. Q.) of standard DNA template (black circles) or target (CXCR4) mRNA (red circles). The plots shown are representative of two similar experiments conducted.