Figure 3. mRNA and protein analyses of IL-6 (A,B,F,G), LIF (C,D,H,I), and IL-11 (E,J) expression in unstimulated controls and fibroblasts stimulated by IL-1β (10- 300 pg/mL) (A,C,E,F,H,J) and TNF- (0.3–30 ng/mL) (B,D,G,I) for 24 hrs (mRNA) and 48 hrs (protein). mRNA levels were assessed by q-PCR and normalized to those of RPL13A, and the values shown are expressed as percent of control, set to 100% (Figs. 3A–3E). Cytokine protein levels in cell lysates and culture media were determined by ELISA. IL-6 protein in cell lysates and LIF and IL-11 protein in culture media were normalized to the total amount of cell protein in cell lysates, measured by the BCA method. Data represent means for 4 wells, and SEMs are shown as vertical bars when larger than the radii of the symbols. Statistically significant (p < 0.05) mRNA and protein stimulation was observed at and above 10 pg/mL (IL-1β) and 0.3 ng/mL (TNF-).