PPARγ activates HUWE1 expression and decreases p65 levels. (A,C-E) HPAECs were transfected with green fluorescent protein (GFP) or AdPPARγ (25 multiplicity of infection) constructs for PPARγ overexpression. After 6 hours, HPAECs were then incubated for an additional 72 hours with DMSO vehicle (CON) or HEM (5 µM). Real-time qPCR was performed for HUWE1 (A,C), miR-98 (D), or p65 (E). Each bar represents mean ± SEM HUWE1, miR-98, or p65 level relative to GAPDH or RNU6B expressed as fold change vs cells treated with GFP. *P < .05 vs GFP or CON/AdPPARγ(−); ⁺P < .05 vs HEM/AdPPARγ(−), n = 5 to 6. (B) Whole lungs were collected from littermate control (FulCon) or endothelial-targeted PPARγ overexpression (ePPARγOX) mice. Levels of lung HUWE1 were measured with real-time qPCR and expressed relative to lung GPADH mRNA. *P < .05 vs FulCon, n = 6. (F-H) The chimeric SS^ePPARγKO mice were generated by transplanting SCD bone marrow from Townes mice using methods we previously published^50-53  into ePPARγKO mice as we have reported.^42,54  ePPARγKO mice were irradiated and transplanted with SS bone marrow. Whole lung homogenates were collected from littermate control (AA^LitCon and SS^LitCon) and chimeric (AA^ePPARγKO and SS^ePPARγKO) mice. Real-time qPCR was performed on lung tissue. Lung HUWE1 (F), miR-98 (G), or p65 (H) levels are expressed relative to 9S or RNU6B normalized to CON values. Each bar represents the mean ± SEM. *P < .05 vs AA^LitCon; ⁺P < .05 vs + SS^LitCon, n = 3 to 7.