TY - JOUR
T1 - Methods for investigating STAT3 regulation of lysosomal function in mammary epithelial cells
AU - Lloyd-Lewis, Bethan
AU - D’Angelo, Michael E.
AU - Prowting, Neve B.
AU - Wiseman, Bethan E.
AU - Sargeant, Timothy J.
AU - Watson, Christine J.
N1 - Publisher Copyright:
© The Author(s) 2024.
PY - 2024/5/18
Y1 - 2024/5/18
N2 - The transcription factor STAT3 is activated by multiple cytokines and other extrinsic factors. It plays a key role in immune and inflammatory responses and, when dysregulated, in tumourigenesis. STAT3 is also an indispensable mediator of the cell death process that occurs during post-lactational regression of the mammary gland, one of the most dramatic examples of physiological cell death in adult mammals. During this involution of the gland, STAT3 powerfully enhances the lysosomal system to efficiently remove superfluous milk-producing mammary epithelial cells via a lysosomal-mediated programmed cell death pathway. The lysosome is a membrane-enclosed cytoplasmic organelle that digests and recycles cellular waste, with an important role as a signalling centre that monitors cellular metabolism. Here, we describe key strategies for investigating the role of STAT3 in regulating lysosomal function using a mammary epithelial cell culture model system. These include protocols for lysosome enrichment and enzyme activity assays, in addition to microscopic analyses of the vesicular compartment in cell lines. Collectively, these approaches provide the tools to investigate multiple aspects of lysosome biogenesis and function, and to define both direct and indirect roles for STAT3.
AB - The transcription factor STAT3 is activated by multiple cytokines and other extrinsic factors. It plays a key role in immune and inflammatory responses and, when dysregulated, in tumourigenesis. STAT3 is also an indispensable mediator of the cell death process that occurs during post-lactational regression of the mammary gland, one of the most dramatic examples of physiological cell death in adult mammals. During this involution of the gland, STAT3 powerfully enhances the lysosomal system to efficiently remove superfluous milk-producing mammary epithelial cells via a lysosomal-mediated programmed cell death pathway. The lysosome is a membrane-enclosed cytoplasmic organelle that digests and recycles cellular waste, with an important role as a signalling centre that monitors cellular metabolism. Here, we describe key strategies for investigating the role of STAT3 in regulating lysosomal function using a mammary epithelial cell culture model system. These include protocols for lysosome enrichment and enzyme activity assays, in addition to microscopic analyses of the vesicular compartment in cell lines. Collectively, these approaches provide the tools to investigate multiple aspects of lysosome biogenesis and function, and to define both direct and indirect roles for STAT3.
KW - Cell death
KW - Lysoptosis
KW - Lysosome
KW - Lysosome-mediated programmed cell death
KW - Mammary gland
KW - STAT3 signalling
UR - http://www.scopus.com/inward/record.url?scp=85193609879&partnerID=8YFLogxK
U2 - 10.1007/s10911-024-09563-3
DO - 10.1007/s10911-024-09563-3
M3 - Article
C2 - 38761238
AN - SCOPUS:85193609879
SN - 1083-3021
VL - 29
JO - Journal of Mammary Gland Biology and Neoplasia
JF - Journal of Mammary Gland Biology and Neoplasia
IS - 1
M1 - 11
ER -