arrayit huprot v2.0 19k human proteome microarray  (CDI Laboratories)

Journal: Cell Death & Disease

Article Title: Mitochondrial Drp1 recognizes and induces excessive mPTP opening after hypoxia through BAX-PiC and LRRK2-HK2

doi: 10.1038/s41419-021-04343-x

Figure Lengend Snippet: A Human proteome microarray showing potential interacting proteins with Biotin-tagged Drp1 protein. Red arrow indicated the positive control (Biotin-BSA), yellow arrow indicated the negative control (BSA protein), and white arrow indicated LRRK2, our interested potential interacting proteins with Biotin-tagged Drp1 protein. B Co-IP results for the combination of Drp1 and LRRK2 in total, cytoplasmic and mitochondrial fractions of VSMCs after hypoxia. ( n = 3 samples/group). C Representative immunofluorescence images showing the co-location of Drp1 (red), LRRK2 (green) and mitochondria (white) in VSMCs after hypoxia. (bar, 5 μm). D Molecular docking of Drp1 to LRRK2 in hypoxia conditions determined by GRAMM-X Protein-Protein Docking program. The blue dotted indicated the stable hydrogen bond between Drp1 Thr595 and LRRK2 Gly2019. E Co-IP results for the combination of Drp1 and LRRK2 in hypoxia-induced VSMCs after Drp1 T595A mutation. F The phosphorylation of HK2-Thr 473 and HK2 expression in total, cytoplasmic and mitochondrial fractions of hypoxia-induced VSMCs after Drp1 T595A mutation. β-actin, Tubulin and COX IV were used as interior references of total, cytoplasmic and mitochondrial fractions. G Representative confocal images of mPTP opening in hypoxia-induced VSMCs after Drp1 T595A mutation. (bar, 100 μm). a: P < 0.05 compared with Normal group.

Article Snippet: Arrayit HuProt™ v2.0 19K Human Proteome Microarrays (CDI Laboratories, Baltimore, MD, USA) were used to identify Drp1-interacting proteins.

Techniques: Microarray, Positive Control, Negative Control, Co-Immunoprecipitation Assay, Immunofluorescence, Mutagenesis, Expressing