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. 2019 Jan 30;196(2):276–286. doi: 10.1111/cei.13259

Figure 5.

Figure 5

Vasoactive intestinal peptide (VIP) prevents tristetraprolin (TTP) from binding interleukin (IL)‐10 mRNA in monocytes (Mos). CD14+CD16– Mos were prepared from blood samples of healthy control (HC) subjects. (a–c) Mos were stimulated with lipopolysaccharide (LPS) for 30 min in the culture, washed and cultured with fresh medium for 48 h. The cells were analyzed by reverse transcription–quantitative polymerase chain reaction (RT–qPCR) and Western blotting. Bars indicate levels of tristetraprolin (TTP) mRNA (a); immunoblots indicate TTP protein levels (b). (c) A portion of the Mos were further treated with the procedures denoted on the x‐axis. Bars indicate IL‐10 mRNA levels in Mos. (d) TTP RNAi results in Mos. (e) Mos were treated with LPS and VIP in the culture for 48 h. The cells were analyzed by immunoprecipitation (IP). Immunoblots indicate a complex of VIP and TTP in Mos. (f) Mos were exposed to LPS or LPS/VIP in the culture for 48 h. The cells were analyzed by RT–qPCR and Western blotting. Bars indicate IL‐10 mRNA (f) and immunoblots indicate TTP protein in a complex in Mos (the β‐actin is from cytosolic extracts serving as a reference). Data of bars are presented as mean ± standard error of the mean (s.e.m.). Each dot inside bars presents data from an independent experiment.