Fig. 3: Interaction between ACL1 and ROC1-8 proteins and ACL1 competition with ROC4/ ROC5 in the formation of homo-dimers and hetero-dimers.

a LCI assay to detect the interaction between ACL1 and ROC1-8 proteins as indicated, respectively. b Co-IP assay to detect the interaction between ACL1 and ROC1-8 proteins as indicated respectively. 1–8, Co-IP of ACL1 and ROC1, ROC2, ROC3, ROC4, ROC5, ROC6, ROC7 and ROC8, respectively. c LCI assay indicating competition of ACL1 with ROC4–ROC4 homodimerization. d Relative LUC activity (LUC/REN) in (c). e LCI assay indicating competition of ACL1 with ROC4–ROC5 heterodimerization. f Relative LUC activity (LUC/REN) in (e). g LCI assay indicating competition of ACL1 with ROC5–ROC5 homodimerization. h Relative LUC activity (LUC/REN) in (g). Data in (d), (f), and (h) are means ± SD (n = 5), and the P-values were determined by a two-tailed unpaired Student’s t test. i–k Co-IP assay showing competition of ACL1 with ROC4–ROC4 homodimerization (i), ROC4–ROC5 heterodimerization (j) and ROC5–ROC5 homodimerization (k). In (i), (j), and (k), the three left lanes showed the protein immunoblots of input controls with anti-HA, anti-GFP, and anti-Flag antibodies. The right three lanes showed the results of Co-IP. The Flag-Trap was used for the co-immunoprecipitation. The experiments in (b) and (i–k) were repeated at least three times with similar results. Source data are provided as a Source Data file.