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Then, the slides were incubated with mouse primary Nrf2 antibody (Proteintech, 66504-1-lg) and rabbit primary PKM2 antibody (Cell Signaling Technology, 4053) at 4 C overnight and then with PLA probe solution for 1 h at 37 C

Then, thrust the needle in like throwing a dart, at a 90 angle (or 45 if you have less fat)