Fig 4.

Heat map of TLR4-dependent gene expression changes induced by cotreatment with palmitate and mmLDL (A). J774 cells were stimulated with palmitate for 16h and incubated with or without mmLDL and LPS. Profiles of mRNA were determined by RNA-seq analysis (B-top). To validate RNA-seq results, independent real time PCRs were used to assess the expression of Ccr5, Il-6, Csf-3, Il-1β, and β–actin (B-bottom). The combination of palmitate and mmLDL caused an increase of mRNA expression of Il-6, Csf-3, and Il-1β genes (normalized to that of Actb). The real time PCR was conducted with technical duplicates, and the data shown represent three independent replicate experiments. *p <0.05 and **p <0.01 compared to LPS treatment without palmitate or mmLDL.