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. Author manuscript; available in PMC: 2022 Sep 14.
Published in final edited form as: Immunity. 2021 Jul 27;54(9):2072–2088.e7. doi: 10.1016/j.immuni.2021.07.003

Figure 6. TRPV4 regulates growth factor expression in macrophages.

Figure 6.

A, mRNA expression of TRP channels in CCR2− macrophages. n=20 samples. B, Ratiometric calcium assay demonstrating that cardiac macrophages have active TRVP4 channels. GSK101: TRPV4 agonist, GSK219: TRPV4 antagonist. * denotes p<0.05 (ANOVA, Post-hoc Tukey) comparing GSK101 treated cells with GSK101 and GSK219 treated cells. C, Immunostaining of Trpv4-GFP BAC transgenic mice showing GFP (green) expression in CD68+ macrophages (red). Scale bar: 20μm. D, Flow cytometry of cardiac CD45+GFP+ leukocytes isolated from Trpv4-GFP hearts. n=4 per group. E, 2-photon imaging of GFP (green) and tdTomato (red) in papillary muscle preparations harvested from Cx3cr1-ertCre; Rosa26-GCaMP6/tdTomato mice treated with either vehicle or the TRPV4 inhibitor GSK219 (TRPV4i). Scale bar: 10μm. F, Quantification of GCaMP6 signal. n=6 per group. * denotes p<0.05 (paired t-test) compared to vehicle. G, Ratiometric calcium assay showing that bone marrow derived macrophages express active TRPV4. GSK101: TRPV4 agonist, Ionomycin: calcium ionophore. H, Cyclic uniaxial stretch (1 Hz, 10% deformation, 24 hours) promotes elongation of bone marrow derived macrophages. n=4 independent experiments. Scale bar: 50μm. * denotes p<0.05 (ANOVA, Post-hoc Tukey) compared to vehicle unstretched. I, Quantitative RT-PCR measuring Igf1, Hbegf, and Cyr61 mRNA expression in stretched macrophages treated with vehicle or TRPV4 inhibitor. n=4 independent experiments. * denotes p<0.05 compared to vehicle treated unstretched cells (ANOVA post-hoc Tukey). J, Quantitative RT-PCR measuring Igf1, Hbegf, and Cyr61 mRNA expression in stretched control, Myd88−/−, and Trif−/− macrophages. n=4 independent experiments. * denotes p<0.05 compared to vehicle treated unstretched cells (ANOVA post-hoc Tukey) (F-H). K, Quantitative RT-PCR of macrophages stimulated with vehicle control, LPS, or polyIC. * denote p<0.05 (Mann-Whitney). n=4 independent experiments. See also Figure S6, Data S2.