Briefly, slides were baked at 65 °C overnight and then dewaxed with Xylene, rehydrated with ethanol gradients and treated with 3% hydrogen peroxide solution for 10 min to inhibit endogenous peroxidase (PO). The slides was immersed in fresh citrate buffer and pressure cooked for 3 min to achieve antigen retrieval. Slides were then incubated with CD147 (1:150, Abcam, USA) and HSP60 (1:300, Santa Cruz, USA) at 4 °C in a wet box overnight. The slides were brought to room temperature (RT) for 30 min on the next day, incubated with universal secondary antibody (anti-mouth/rabbit, 1:200, Zhongshan Company, Beijing, China) for 1 h. Slides were treated with DAB (1:100, Zhongshan Company, Beijing, China) 15 s and then stained with Hematoxylin.
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