10.1371/journal.pone.0096286.g005 Jack H. Wang Jack H. Wang Kosei Eguchi Kosei Eguchi Sahohime Matsumoto Sahohime Matsumoto Katsuhito Fujiu Katsuhito Fujiu Issei Komuro Issei Komuro Ryozo Nagai Ryozo Nagai Ichiro Manabe Ichiro Manabe EPA reduces <i>Mmp9</i> expression in macrophages. Public Library of Science 2014 anatomy Cardiovascular anatomy Biochemistry lipids Fatty acids cell biology Cellular types Animal cells Blood cells White blood cells monocytes Immune cells immunology cardiology Cardiovascular diseases Valvular diseases Diagnostic medicine Diagnostic radiology tomography Computed axial tomography Radiology and imaging Vascular medicine Aortic diseases Model organisms Animal models Mouse models 2014-05-05 03:30:16 Figure https://plos.figshare.com/articles/figure/_EPA_reduces_Mmp9_expression_in_macrophages_/1016205 <p><b>A</b>. Gelatin zymography of aortic tissues one week after CaCl<sub>2</sub> treatment together with quantitative analysis, showing reduced MMP9 activity in samples from the EPA diet group. Equal amounts of protein (20 µg) were loaded per aortic sample. For quantitation, <i>n</i> = 6–7 in each group. <b>B</b>. Gating strategy for the flow cytometric analysis of AAA macrophages. Macrophages were identified as Ly-6C<sup>low</sup>CD11b<sup>+</sup>F4/80<sup>+</sup>Ly-6G<sup>−</sup> cells (full gating strategy shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096286#pone.0096286.s001" target="_blank">Figure S2 in File S1</a>). <b>C</b>. The number of aortic macrophages per aortic sample. No statistically significant difference in the number of aortic macrophages between control diet and EPA diet groups was detected. <b>D</b>. The mRNA levels of <i>Mmp9</i> in sorted aortic macrophages. Expression levels were first normalized to <i>18s</i> rRNA levels and then further normalized to the level of control diet group. <i>n</i> = 5 in each group. <b>E</b>. RAW264.7 macrophages were cultured with either vehicle (10% BSA) or EPA (50 µmol/L) for 48 hours. The cells were then stimulated with recombinant mouse TNF-α (20 ng/mL) for a further 6 hours and harvested for analysis by RT-PCR. Expression levels were first normalized to <i>18s</i> rRNA levels and then presented as relative expression compared to baseline vehicle sample. <i>n</i> = 3 per condition. *<i>P</i><0.05 compared to control diet group in <b>A</b> and <b>D</b> or respective vehicle controls in <b>E</b>.</p>