Ab215166

Mouse tumor samples were fixed in 4% paraformaldehyde and embedded in paraffin. Tissue slices were deparaffinized, rehydrated, and antigen retrieval was performed using 10 mM citrate buffer (pH 6.0). CD80 (polyclonal rabbit, 1:200, ab215166, Abcam), CD86 (monoclonal rabbit, 1:200, ab243887, Abcam), and CD69 (polyclonal rabbit, 1:250, A00529-2, Boster) were stained overnight at 4 °C. The samples were washed and then incubated with goat anti-rabbit biotinylated secondary antibody (1:1000, ab6720, Abcam) and visualized using a horseradish peroxidase (HRP)-conjugated ABC system (1:10,000, ab7403, Abcam).

The antibodies used were as follows: rabbit anti-RhoA (1 : 300, Abcam, ab187027), rabbit anti-ROCK (1 : 200, Abcam, ab125025), rabbit anti-CD 68 (1 : 200, Abcam, ab125212), rabbit anti-CD80 (1 : 200, Abcam, ab215166), rabbit anti-CD163 (1 : 200, Abcam, ab182422), rabbit anti-CD206 (1 : 200, Abcam, ab64693), rabbit anti-inducible nitric oxide synthase (iNOS, 1 : 200, Abcam, ab178945), mouse antitumor necrosis factor-α (TNF-α, 1 : 200, Santa Cruz, sc-52746), mouse anti-interleukin-6 (IL-6, 1 : 300, Abcam, ab9324), and mouse anti-IL-10 (1 : 200, Santa Cruz, sc-365858).