Autophagy and Lipid Signaling Pathway Protocols

The following rabbit antibodies were used: Atg16L1 (#PA1-18296 Thermo Scientific), beclin-1 (#sc-11427 Santa Cruz), and LC3 (#L7543 Sigma, #PM036 MBL). The following mouse antibodies were used: beta-actin (#sc-81178 Santa Cruz), galectin-3 (#556904 BD Pharmingen), and ubiquitin (FK2, #BML-PW8810 Enzo Life Sciences). Pam3CSK4 was purchased from Invivogen. The synthetic SL analog (2,3-dipalmitoyl-2′-sulfate-α-α′-d-trehalose) we used was a kind gift from Drs. Jacques Prandi and Martine Gilleron (Institute of Pharmacology and Structural Biology, Toulouse) [28 (link),29 (link)]. The DIMs were prepared as previously described [30 (link)].

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Bah A., Sanicas M., Nigou J., Guilhot C., Astarie-Dequeker C, & Vergne I. (2020). The Lipid Virulence Factors of Mycobacterium tuberculosis Exert Multilayered Control over Autophagy-Related Pathways in Infected Human Macrophages. Cells, 9(3), 666.

Publication 2020

Atg16L1 beclin-1 beta-actin galectin-3 ubiquitin (FK2 Pam3CSK4

Antibodies Beclin 1 Beta actin Dims Galectin 3 Mouse Rabbit Sulfate Trehalose Ubiquitin

Corresponding Organization :

Other organizations : Institut de Pharmacologie et de Biologie Structurale, Université Claude Bernard Lyon 1

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Variable analysis

independent variables

Pam3CSK4
Synthetic SL analog (2,3-dipalmitoyl-2′-sulfate-α-α′-d-trehalose)

dependent variables

Expression of Atg16L1, beclin-1, and LC3 proteins
Expression of beta-actin, galectin-3, and ubiquitin proteins

control variables

Primary antibodies used for Western blotting (Atg16L1, beclin-1, LC3, beta-actin, galectin-3, ubiquitin)

controls

Positive control: None specified
Negative control: None specified

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