Fig. 3: CaLB1 interacts with ATG8. | Nature Communications

Fig. 3: CaLB1 interacts with ATG8.

From: Arabidopsis CaLB1 undergoes phase separation with the ESCRT protein ALIX and modulates autophagosome maturation

Fig. 3

a In vitro binding assays of GST and CaLB1-GST with His-tagged ATG8s bound to TALON beads. Bound proteins were analyzed with an anti-GST antibody. His-ATG8s were visualized by protein staining using CBB. The experiment was conducted twice, and one representative result is shown. b MicroScale Thermophoresis analysis of the C2 ___domain of CaLB1 with ATG8i or ubiquitin (UB). The changes in fluorescence are shown in relation to the concentration of ATG8i or UB. The experiment was conducted twice, each time with three technical replicates. One representative result is shown. Error bars: standard deviation. c Yeast two-hybrid analysis of the regions in CaLB1 responsible for the interaction with ATG8i. Four aromatic amino acids (Y34, F64, Y172, and Y205, positions shown in the scheme) were each mutated and the impact of the mutation was tested in a yeast two-hybrid assay. PRD proline-rich ___domain, -LW growth media lacking leucine and tryptophan, -LWH growth media lacking leucine, tryptophan, and histidine, 3-AT 3-Amino-1,2,4-triazole. The experiment was conducted three times, and one representative result is shown. a, b Source data are provided as a Source Data file.

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