Figure 4. R9-SOCS3-KIR suppressed LPS-induced nuclear translocation of NF-κB and MAP kinase p38 in J774A.1 cells. The J774A.1 cells were seeded on an eight-well cell culture slide and grown overnight. The cells were transferred to a serum-free medium and pre-treated with R9-SOCS3-KIR peptide or its control peptide at 20 μM for 1 h, followed by treatment with LPS (1μg/ml) for 30 min. The cells were stained with an antibody to p65, which is the active subunit of NF-κB (A), or phosphorylated p38 (p-p38; B). Secondary antibodies conjugated to Alexa-488 (A, green) or Cy3 red (B, red) and DAPI (nuclei, bottom row in A and B) were used and imaged using a fluorescence microscope. Scale bar: 50 μm.