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. Author manuscript; available in PMC: 2020 Jul 1.
Published in final edited form as: Mol Cancer Res. 2019 Oct 16;18(1):166–178. doi: 10.1158/1541-7786.MCR-19-0204

Figure 7: PRMT6 overexpression promotes alternate activation of tumor-associated macrophages.

Figure 7:

A. Lung tumor sections of PRMT6Tg; Sftpc-Cretm and PRMT6Tg control mice after tamoxifen and urethane treatment (20 weeks) were subjected to indirect immunofluorescence staining with anti-CD68, anti-iNOS, and anti-arginase I antibodies as described in the methods. Scale bar: 20 μM. B. Gating strategy for the purification of TAMs using CD11b antibodies. Sixty% of live cells are CD11b+, of which 83% are F4/80+. C. Total RNA from CD11b+ cells from the lungs of PRMT6Tg; Sftpc-Cretm and PRMT6Tg control mice after tamoxifen and urethane treatment (20 weeks) were employed in qPCR analysis for the indicated M1 and M2 macrophage markers. *, p< 0.05; versus control. D. Bone marrow-derived macrophages were treated with the conditioned medium isolated from AECs isolated from PRMT6Tg; Sftpc-Cretm or PRMT6Tg control mice after tamoxifen treatment. Later, total RNA from the macrophages were isolated and qPCRs were performed for M1 and M2 macrophage markers as described in the methods. **, p< 0.01; *, p< 0.05; versus control. E. THP1 cells were incubated with 150 mM phorbol myristate acetate (PMA) for 24h. Later, the differentiated macrophages were incubated with conditioned medium from H2122 PRMT6 knockout clone or parental cells for additional 24 h. Total RNA from the macrophages were isolated and qPCRs were performed for M2 macrophage marker as described in the methods. **, p< 0.01. F. Total RNA isolated from bone marrow-derived macrophages treated with the conditioned medium isolated from AECs isolated from PRMT6Tg; Sftpc-Cretm or PRMT6Tg control mice after tamoxifen treatment were employed in qPCRs for pro-angiogenic gene markers as described in the methods. *, p< 0.05; versus control. G. Total RNA isolated from lungs of PRMT6Tg; Sftpc-Cretm and PRMT6Tg control mice after tamoxifen and urethane treatment (20 weeks) were subjected to qPCR analysis of pro-angiogenic gene markers as described in the methods. **, p< 0.01 *, p< 0.05; versus control. H. Lung tumor sections of PRMT6Tg; Sftpc-Cretm and PRMT6Tg control mice after tamoxifen and urethane treatment (20 weeks) were subjected to indirect immunofluorescence staining with anti-CD31 antibodies as described in the methods. Scale bar: 10 μM.