TREM-1 activation instigated glucose metabolic reprogramming in macrophages. Macrophages were stimulated with 10 μg/mL of an agonist anti-TREM-1 mAb. (A) Six hours later, the lactate in the supernatant was assayed, n=3. (B) Glucose consumption in the supernatant in control or 24 h TREM-1-activated macrophages, n=3. (C)
Glut-1, Hk2, Pfkfb3, Pkm2, Ldha, and Hif-1α mRNA in control or 6 h TREM-1-activated macrophages. Data was normalized to housekeeping gene β-actin, n=3-5. (D-F) Western blot and quantification of the glycolytic enzymes (HK2 and LDHA) in the control or 24 h TREM-1-activated macrophages, n=3. (G) OXPHOS-related proteins: ATP5A, MTCO1, UQCRC2, SDHB, and NDUFB8, in the macrophages were detected by western blot. (H-J) Quantification of indicated protein levels in (G), n=3. Macrophages were administrated with LR12 (25 μg/mL) 30 min before the LPS stimulation (1 ng/mL). Twenty-four hours later, (K) the lactate and (L) relative glucose consumption in the supernatant were assayed, n=3. (M) Expression of Glut-1, Ldha, and Hif-1α mRNA in macrophages was detected by real-time PCR, n=3. n represents experiments performed on different macrophages from separate mice. Bar graphs represent mean ± SD. Student's t-test (two-tailed, unpaired) was used to compare Mab1187 and Control in (A-J): * P < 0.05, ** P < 0.01, and *** P < 0.001. One-way ANOVA adjusted by Tukey's multiple comparison test was used in (K-M): * P < 0.05, ** P < 0.01, and *** P < 0.001.