Fig. 4.

CaMKK2 regulates gene transcription in BMDM. Bone marrow-derived macrophages (BMDM) were generated from WT and Camkk2^−/− mice in the presence or absence of E0771-conditioned medium (RM and TCM, respectively). Subsequently, BMDM was harvested and analyzed for gene expression. N = 4 biological replicates/group. a, b Volcano plots of differentially expressed genes (DEGs) in WT (left) and Camkk2^−/− (right) generated in the presence of RM or TCM. Open circles indicate genes showing comparable expression levels in WT and Camkk2^−/− BMDM. DEGs between WT and Camkk2^−/− BMDM are shown as green filled circles. Yellow open circles refer to the Camkk2 gene. c Heatmaps of DEGs affiliated with M1, M1 and dendritic cells (M1&DC), or M2 signatures. The color key for the heatmap indicates (row-wise) scaled RPKM values (z-score). d Real-time quantitative PCR (qPCR) analysis of genes associated with M1 (Nos2) and M2 (Arg1, Chil3/Ym1 and Retnla/Fizz1) gene signatures (four biological replicates were analyzed for each genotype). A t test was used to calculate p-values. The data are graphed as the mean ± SEM. Asterisks refer to *p < 0.05, **p < 0.01, ***p < 0.005, and ****p < 0.001. This experiment was replicated at least three times with similar results