Fig 1: (A) mRNA levels of enzymes involved in nucleotide metabolism in mouse liver, Actb was chose as the housekeeping genes to normalize the qPCR results. (B) Protein levels of ADSL, APRT, PNP, and ADA in HepG2 cells after serum shock. (C) Urate levels in cell culture supernatant at different points after serum shock. (D) Per2 mRNA levels in liver after T3 treatment, Actb was chose as the housekeeping genes to normalize the qPCR results. (E) Per2 mRNA levels in HepG2 cells at different time points after serum shock, Actb was chose as the housekeeping genes to normalize the qPCR results. (F) Protein levels at 48 h after transfection with siPer2. (G) Per2 levels in HepG2 cells after serum shock. (H) Urate production in HepG2 cells at 48 h after treatment (*P < 0.05; **P < 0.01).
Fig 2: TREM2+ TAMs display purine metabolic activity and are immunosuppressive(A) Western blotting of APRT and PNP in F4/80+ cells sorted from liver with and without tumor.(B and C) Metabolic activity of PNP. PNP was detected in TREM2+ TAMs and hepatic F4/80+ macrophages from normal liver. Student’s t test was used to calculate the significance between two groups.(D and E) Levels of key purine pathway metabolites. AMP (D) and adenosine (E) were measured in TREM2+ TAMs and hepatic F4/80+ macrophages from normal liver. Student’s t test was used to calculate the significance between two groups.(F–H) Flow cytometry quantification of TREM2+ hepatic macrophages in liver with and without tumor. Percentage of TREM2+ hepatic macrophages in CD11b+F4/80+ macrophages (G) and cell number of TREM2+ hepatic macrophages (H) are shown. Student’s t test was used to calculate the significance between two groups.(I and J) Flow cytometry analysis of MHC-II (I-A/I-E) (I) and MHC-I (H-2Db) (J) expression in TREM2+ TAMs and macrophages from liver without tumor. Student t-test was used to calculate the significance between two groups.(K and L) Flow cytometry analysis of OT-I cell proliferation in the presence or absence of TREM2+ and TREM2− cells purified from liver with tumor. Representative histogram (K) and the percentage of OT-I cells at the third generation (L) are shown. The ratio of TREM2+ or TREM2− cells to OT-I cells is 4:1.
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