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. 2018 Nov 27;9:1386. doi: 10.3389/fphar.2018.01386

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Copyright © 2018 Molteni, Rizzi, Bresciani, Meanti, Fehrentz, Verdié, Omeljaniuk, Biagini, Locatelli and Torsello.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Effect of C3aR siRNA on RAW264.7 cells stimulated with JMV5656 and C3a_(70-77). Graphs show mRNA (A) and protein (B) levels of C3aR after transfection with 25 mM siRNA for 24 h. Data are shown as the mean ± SEM of measurements obtained in at least three independent experiments. ^∗∗p < 0.01, ^∗∗∗p < 0.001 vs. control (CTRL). (C) After 24 h from transfection with 25 mM siRNA, cells were loaded with FLUO-4 NW and fluorescence emissions were measured at 485/535 nm every 0.5 s for the 20 s preceding and the 60 s following the injection of the stimuli. Graphs show intracellular Ca²⁺ levels (expressed as fluorescence intensity) in RAW264.7 cells stimulated with HBSS (vehicle), JMV5656, and C3a_(70-77) in the absence or presence of C3aR siRNA. Results are shown as the mean ± SEM of measurements obtained in three independent experiments (n = 18). ^∗p < 0.05 vs. C3a CTRL (Student’s t-test).