Fig. 5: In vivo studies of effects of lentivirus carrying CD11b promoter upstream of Cre recombinase to silence Tgm2 in TGM2fl/fl male mice.
From: Transglutaminase 2-expressing macrophages modulate adipose tissue inflammation
Tgm2 floxed mice (B6.129S1-Tgm2tm1Rmgr/J) were injected at 5 weeks of age with A pLV.Control or pLV.CD11b-Cre lentiviruses, followed by CD or 60% HFD treatment and further metabolic evaluations. B Timeline for procedures. C Flow cytometric gating strategy employed for eWAT-derived SVF from pLV.Control vs. pLV.CD11b-Cre treated mice and D subsequent analysis looking at % of TGM2+ATMs (CD45+ CD11b+GFP+ F4/80+ MHCII+), Leukocytes (CD45+), MHCIIlo+ATMs (CD45+ CD11b+GFP+ F4/80+ MHCII-), MHCIIhi+ATMs (CD45+ CD11b+GFP+ F4/80+MHCIIhi+), IL-10+ATMs (CD45+ CD11b+GFP+ F4/80+ MHCII+ IL-10+ ), and AT T cells (CD45+ CD11b+GFP+ F4/80-MHCII-TCRb+) (n = 5/group). E LegendPlex assessment of cytokine inflammatory profile in serum from pLV.Control vs. pLV.CD11b-Cre injected mice (n = 5/group). Data are shown as bar graphs with SEM of five mice per control or treated group and are representative of two independent experiments. Graphs were statistically analyzed by Two-way ANOVA for flow cytometric populations. For all bar graphs, data normality status was used to determine statistical analysis. Normal data statistical significance was determined by parametric student’s unpaired t test, while not normally distributed data were analyzed using non-parametric Mann-Whitney test. ns = P > 0.05, *=P < 0.05, ** =P < 0.01, ***P < 0.001.
