Stat1-induced NAMPT sustains glycolysis to regulate gene expression in classically activated macrophages

Single Cell RNA Sequencing is used to profile in vivo responses by immune cells lacking the Nampt NRE1 region to melanoma challenge, finding multiple defects in macrophages and other myeloid APCs. We analyzed B16F10 melanoma tumor infiltrating immune cells from mice specifically lacking NRE1 in hematopoietic cells, finding inflammatory and metabolic defects in APC populations within tumors, correlating with trends toward defective control of malignant growth. Overall design: 500,000 B16F10 cells (from Mingnan Chen lab) injected subcutaneously into NRE1 fl/fl VavCre(-) or NRE1 fl/fl VavCre(+) male mice. Tumors were excised at the endpoint, pooled and digested with Accumax to liberate single cells. Cells were stained with APC-CD45 (1:200) for 15 min on ice and sorted via flow cytometer (HCI Aria) after spiking 1:100 DAPI (1x final). CD45+DAPI- cells were obtained (860,000 cells from WT, 1,0260,00 cels from KO), and subjected to Single Cell RNA Sequencing.