Yeloid-Specific PHD2-Deficient MicePHD2 expression in different organs from control and MyPHD2KO mice had been examined by Western blot analysis (Figure 1A). PHD2 protein was not detected in BMDMs and PMs from MyPHD2KO mice. Phd2 mRNA expression determined by RT-qPCR in BMDMs and PMs from MyPHD2KO miceJournal of your American Heart AssociationAttenuation of Cardiovascular Remodeling by Phd2 DeletionIkeda et alORIGINAL RESEARCHAPHD2 -Tubulin C KO C KO PM BMDM Phd2 / Hprt mRNA C KO C KO C KO C KO 46kDa 50kDaBMDMHeartLung120 100 80 60 40 20LiverKidney PMBPhd2 / Hprt mRNA120 100 80 60 40 20**Control MyPHD2KO n.s.**Control MyPHD2KOPhd1 / Hprt mRNAC1.Phd3 / Hprt mRNA20 15 ten 5**0.Control MyPHD2KOControl MyPHD2KODHIF-1 HIF-2 Histone H3 100kDa 118kDa 17kDa Control MyPHD2KOEHIF-1 / Histone H10 eight six four 2F **HIF-2 / Histone H3 two 1**Control MyPHD2KOControl MyPHD2KOFigure 1. Suppression of PHD2 protein and mRNA expression in MyPHD2KO mice. A, Western blot for PHD2 in bone marrow derived macrophages (BMDMs), peritoneal macrophages (PMs), heart, lung, liver, and kidney in handle and MyPHD2KO mice is shown. Precisely the same results were obtained in other 2 independent experiments. B, Phd2 mRNA expression in BMDMs and PMs was measured by RT-qPCR. **P0.01 vs Handle. n=8. C, Phd1 and Phd3 mRNA expression in PMs were measured by RT-qPCR. **P0.01 vs Control. n=8. D, Western blot for HIF-1a and HIF-2a in nuclear protein of PMs is shown. E and F, The bar graphs indicate the expression ratio of HIF-1a and HIF-2a to Histone H3, respectively. **P0.01 vs Handle. n=3. PHD2 indicates prolyl hydroxylase domain protein 2; C, Control; KO, MyPHD2KO; RT-qPCR, real-time reverse transcription-quantitative polymerase chain reaction; HIF, hypoxia-inducible factor; Hprt, hypoxanthine phosphoribosyl-transferase.was substantially suppressed compared with handle mice (Figure 1B). The expression of Phd2 mRNA within the aorta was comparable in between manage and MyPHD2KO mice (information not shown). Next we examined the expression of other isoforms of Phd in PMs.Apiin supplier Though Phd1 mRNA level was unchanged, mRNA of Phd3, a target gene of HIF-1a,20 was drastically larger in PHD2-deficient PMs (Figure 1C).Prostratin manufacturer HIF-1a and HIF-2a protein levels had been substantially enhanced in PHD2-deficient PMs compared with handle PMs (Figure 1D via 1F).PMID:23290930 Expression of HIF-1a and HIF-2a protein was also increased inDOI: 10.1161/JAHA.113.BMDMs from MyPHD2KO mice (information not shown). No considerable adjustments had been noted on full blood cell count in between handle and MyPHD2KO mice (Table 2).Phd2 Deletion Decreased the Expression of M1 Macrophage Markers in PMsTo investigate no matter if PHD2 regulates macrophage polarization, we examined the expression of M1 and M2 macrophage markers in isolated PMs. The expression of Tnfa, Il6, Il1b,Journal on the American Heart AssociationAttenuation of Cardiovascular Remodeling by Phd2 DeletionIkeda et alORIGINAL RESEARCHTable 2. Comprehensive Blood Cell CountControl MyPHD2KO Control+L/A MyPHD2KO+L/A MyPHD2KO+L/A+DigoxinWBC, 910 /lL116.61.4 910.84.1 12.five.four 78.7.117.60.1 870.08.six 12.0.4 72.eight.107.47.3 800.40.0 11.five.9 68.five.117.65.4 915.06.1 12.7.0 70.1.97.04.7 892.00.1 12.5.5 71.eight.RBC, 910 /lLHb, g/dL PLT, 9104/lLData are expressed as mean EM. n=5. L/A indicates L-NAME+Angiotensin II; WBC, white blood cell; RBC, red blood cell; Hb, hemoglobin; PLT, platelet; SEM, normal error of the mean.Rantes, Mcp1, and iNOS, markers of M1 macrophage, was considerably lowered by Phd2 deletion (Figure 2A). Having said that M2 macrop.
