Anti cd86

BV-2 cells were purchased from the Cell Bank of the Chinese Academy of Sciences. Lipopolysaccharide (LPS) was purchased from Sigma−Aldrich (St. Louis, MO, USA). Chlorogenic acid was purchased from Shanghai Yuanye Biotechnology Co. Ltd. (Shanghai, China). Anti-iNOS (1:1000, ab178945) and anti-MMP9 (1:1000, ab228402) were purchased from Abcam. Anti-CD86 (1:1000, BM4121), anti-Arg-1 (1:1000, M01106-4), anti-IL-10 (1:1000, RP1015), anti-CD206 (1:1000, A02285-2), anti-CXCL12 (1:1000, BA1389), anti-CXCR4 (1:1000, A00031-4), anti-PTGS2 (1:1000, A00084), and anti-TNF (1:1000, BA0131) were purchased from Boster Biological Technology Co. Ltd. (Shanghai, China). Cleaved caspase-3 (1:500, GB11532), IBA-1 (1:500, GB12105), and CD206 (1:400, GB113497) were purchased from Wuhan Servicebio Biotechnology Co. Ltd. (Wuhan, China). p-Akt1 (1:1000, 9018S), Akt1 (1:1000, 75692S), p-NF-κB (1:1000, 3033S), NF-κB (1:1000, 8242S), p-ERK1/2 (1:1000, 4370T), ERK1/2 (1:1000, 4696S), p-P38 (1:1000, 4511T), and P38 (1:1000, 8690T) antibodies were purchased from CST.

IHC staining was performed on 5-μm sections of paraffin-embedded tissue samples to detect the protein expression levels of CD68, CD86, CD163, CPEB3, E-cadherin, vimentin and Ki67. In brief, the slides were incubated in anti-CD68(1:500, Servicebio, Wuhan, China), anti-CD86 (1:100, BOSTER, Wuhan, China), anti-CD163 (1:500, BOSTER), anti-CPEB3 (1:200, Abcam, Cambridge, MA, USA), anti-E-cadherin (1:1000, Proteintech, Chicago, IL, USA), vimentin (1:1000, Proteintech) and Ki67 (1:1000, Proteintech) antibodies at 4 °C overnight. All slides were independently evaluated by two observers. The score for CPEB3, E-cadherin and vimentin staining was based on the integrated staining intensity and the proportion of positive cells. IHC staining of CD68, CD86, CD163 and Ki67 was calculated by the positive cell numbers in the per high field. All the percentages/numbers of positive cells were expressed as the average of six randomly selected microscopic fields.