Figure 3.

SPIC as a candidate transcriptional regulator of macrophage specification in fasting adipose tissue. A) Tukey box plot of SPIC expression in human peripheral blood monocytes treated for 24 h with hemin or fatty acids. ∗p < 0.05, ANOVA with Dunnett's multiple comparisons test (n = 4 technical replicates, similar induction to fatty acids in 3 independent experiments). B) Transcriptional pattern resulting from SPIC gain of function compared to SPI1 gain of function. Thp1 cells were transduced with lentiviral vectors delivering doxycycline-inducible SPIC. Control and experimental cells were treated with doxycycline for 48 h after viral transduction, then differentiated via 48-hour PMA stimulation and 24-hour rest to induce adherence to the culture dish prior to qPCR analysis. Results from 7 independent experiments are shown as a heat map (top), demonstrating directional concordance of SPIC induction relative to SPI1. Bottom: dot plots, where each dot represents the mean of 4 technical replicates. To compare the scope of transcriptional changes across experiments, each biological replicate was normalized to vector control (blue line at log₂ = 0). Genes associated with M1 or M2 phenotypes are grouped together. Significance assessed by one sample testing: two-sided t test for normally distributed data or Wilcoxon signed-rank test for non-normal datasets (∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)