Journal: Cell Death & Disease

Article Title: LZTR1 deficiency exerts high metastatic potential by enhancing sensitivity to EMT induction and controlling KLHL12-mediated collagen secretion

doi: 10.1038/s41419-023-06072-9

Figure Lengend Snippet: A The results of liquid chromatography-tandem mass spectrometry. B How LZTR1 regulates SEC31A ubiquitination and inhibits collagen secretion through the inhibition of KLHL12 function. C Ascorbate chase analyses were performed by adding ascorbate (0.25 mM ascorbic acid and 1 mM ascorbic acid 2-phosphate) to MEFs from Lztr1 +/+ or Lztr1 −/− mice. Cells were stained with anti-COL1A1 antibody (green) and NucBlue Stain (nuclei, blue). Scale bar, 20 µm. Values are the mean ± SD; ** P ≤ 0.01 (Wilcoxon-Mann-Whitney test, n = 5). D , E HEK293 cells were transfected with the indicated expression plasmids, and the lysates were subjected to co-immunoprecipitation assays using anti-FLAG M2 Magnetic Beads or anti-GFP mAb-Magnetic Beads. The immunoprecipitants were subjected to western blot analyses. F An in vivo ubiquitination assay was performed with anti-HA-tag mAb-Magnetic Agarose. The ubiquitination status of SEC31A was evaluated by western blot.

Article Snippet: The A549 (CRM-CCL-185) and HEK293 (CRL-1573) cell lines were purchased from ATCC (Manassas, VA, USA).

Techniques: Liquid Chromatography, Mass Spectrometry, Inhibition, Staining, MANN-WHITNEY, Transfection, Expressing, Immunoprecipitation, Magnetic Beads, Western Blot, In Vivo, Ubiquitin Assay