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. 2020 Sep 29;3:536. doi: 10.1038/s42003-020-01268-7

Fig. 2. Engrailed 2 intercellular transfer is asymmetrically regulated by H2O2 in HeLa cells.

Fig. 2

a Method for quantification of EN2 secretion synchronized at the plasma membrane via a modified RUSH strategy, the transRUSH method26. EN2 is tagged with SBP (black) that hooks the protein at the inner side of the plasma membrane in presence of a Streptavidin hook (gray) and HiBiT (white bulb) that emits light when combined to LgBit (yellow crescent). b–e Quantification of EN2 secretion from cells expressing Lck-DAO with or without d-Ala b, expressing inactive or active Catalase (CATmut or CAT, respectively) c, treated with extracellular CAT (CAText,) d, or treated with Nox-i e. f Method for quantification of EN2 internalization. g–i Quantification of EN2 internalization in cells expressing Lck-DAO with or without d-Ala g, treated with CAText h or Nox-i i. *p-value ≤ 0.05; **p-value ≤ 0.01; ***p-value ≤ 0.001; and ****p-value ≤ 0.0001.