Extended Data Fig. 5: P2Y11 was identified as the main receptor of HD5 for HIFE generation.
a, Expression analysis of Gs-coupled GPCRs in the tested cell lines using HPA database. b, Transcriptome analysis of the expression of GPCRs in the tested cell lines. c, Mass spectrometry analysis of the biotin-HD5-pulldown proteins using P2Y11-target searching strategy and two peptides of P2Y11 are present in the HD5-associated protein complex. d, 100-ns dynamic trajectory of simulated interaction between HD5 and P2Y11. e, MD simulation showing that NF340 exhibited -2.99 kcal/mol binding free energy to weakly occupy the receptor P2Y11. f-g, Fluorescence microscopy analysis and quantification of influence of NF157 on cytoskeleton of HeLa cells when added after HD5 addition or before HD5 addition. F-actin is red and nuclei are blue (DAPI). The bar represents 20 μm. Data are presented as mean±s.d. Significance between the indicated groups was calculated using a one-way ANOVA (Tukey’ s multiple comparison Test), and p values are as follows: **p < 0.01, ***p < 0.001 and ****p < 0.0001.h, Immunoblotting analysis of P2Y11 expression after CRISPR/Cas9 knockout. All results are representative of at least three independent experiments.
