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Effects of paracrine factors and cell contact on IL-10 secretion and expression.

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posted on 21.10.2013, 03:16 authored by Carolina Franco Nitta, Robert A. Orlando

(A) Differentiated 3T3-L1 adipocytes or wild type murine splenocytes were cultured alone (columns 1 and 2 or 3 and 4, respectively) or together with either no contact (columns 5 and 6) or direct contact (columns 7 and 8). Cells were incubated in the absence (-) (columns 1, 3, 5 and 7) or presence (+) (columns 2, 4, 6 and 8) of LPS (1 µg/mL) for 24 h as indicated. Interleukin-10 (IL-10) in culture media was quantified by capture ELISA. (B) Differentiated 3T3-L1 adipocytes were co-cultured with no contact or direct contact with GFP-expressing murine splenocytes as in Figure 3 and activated by incubation with LPS (1 µg/mL) for 24 h. Splenocytes were sorted as GFP-positive cells by FACS and IL-10 mRNA expression was measured by qRT-PCR. qRT-PCR values were normalized to values obtained for 36B4. In (A) experimental points were measured in triplicate for calculation of means and standard deviations. Comparison between all conditions was calculated using ANOVA followed by the post-hoc Bonferroni test. No statistical significance was found between any measured points (p>0.05). For (B), experimental points were performed in duplicate, and a statistical comparison between no contact and direct contact was made using an unpaired two-tailed Student's t-test.