The markers of canonical TLR signaling, such as phosphorylated P65 (RELA, NFκB subunit) and phosphorylated P38 MAPK, were both elevated in the NEC group but repressed with butyrate pretreatment, although the expression of TLR4 was not altered (Figures 4A,B). This evidence concerns the gene RELA and necrotizing enterocolitis.