Introducing glyoxal largely preserves protein epitopes and fluorescent protein signals. (A) GO/PFA fixation preserves the protein epitopes during immunostaining. Statistics were counted from more than 30 cells across three independent experiments under each condition. Mean ± SD and paired t-test are shown. Scale bar: 5 µm. (B) GO/PFA fixation provides a better colocalization accuracy in immunostaining shown by higher P's colocalization value between the CRISPR/Cas9-mediated EGFP KI at the endogenous RPA194 locus and the antibody labeled RPA194 proteins. Statistics were counted from more than 1500 RPA194 condensates (the FC regions in nucleoli) across three independent experiments under each condition. Mann–Whitney test is shown. Scale bar: 1 µm and 400 nm for zoomed regions. (C) GO/PFA fixation largely preserves the fluorescent protein signals during RNA FISH, represented by the slower decreasing rate of fluorescent protein signals during FISH process under GO/PFA fixation than PFA alone. Statistics were counted from more than 20 cells across three independent experiments under each condition. Mean ± SD and paired t-test are shown. Scale bar: 10 µm.
