Fig. 2

Effects of rolipram on the LPS-induced DUSP1 expression and inflammatory response in cardiac fibroblasts. Cells were pretreated with 10 μmol/L rolipram for 1 h before LPS stimulation. DUSP1 mRNA and protein expression, the mRNA expression and supernatant protein concentrations of inflammatory mediators in cardiac fibroblasts were measured after LPS stimuli for 6 h. For the RNAi experiment, cardiac fibroblasts were treated by small interfering RNA for 6 h, followed by normal culture for 48 h, then cells were pretreated with 10 μmol/L rolipram for 1 h before LPS stimulation. The expression of DUSP1 and the TNF-ɑ, IL-6 concentrations in the supernatant of cardiac fibroblasts were measured. a, Relative fold of changes to control of DUSP1 mRNA; b and c, DUSP1 expression detection and quantification (original, uncropped blot was shown in supplement Fig. S1B). d-f, Relative fold of changes to control of TNF-ɑ, IL-6 and IL-1β mRNA expression. g-i, Concentrations of TNF-ɑ, IL-6 and IL-1β in the supernatant. j. The expression of DUSP1 after RNAi (original, uncropped blot was shown in supplement Fig. S1C). k and l. The TNF-ɑ and IL-6 concentrations in the supernatant after the DUSP1 knockdown* represents P < 0.05