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Fixed cells were treated with 4% paraformaldehyde (PFA), washed with PBS, permeabilized with 0.5% Triton-X 100 (T8787, Sigma) at room temperature for 1 h, and then incubated overnight at 4 with the following primary antibodies: anti-Ki67 (#145698-82, 1:50, Thermofisher) and anti-CD69 (#MA1-207, 1:50, Thermofisher)