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The cell blocks were prepared for optimization and validation of pMLKL staining and in situ PLA assay. HT-29 cells were solidified using iPGell (Genostaff, Tokyo, Japan) according to the manufacturer’s instructions. Briefly, cells were collected and fixed with 10% neutral-buffered formalin for 10 min. Solutions A and B were added to solidify the cells. The clot was gently removed from the tube and transferred to a tissue cassette. Subsequently, a tissue cassette was soaked into ethanol and xylene and finally embedded in paraffin.
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