TY - JOUR T1 - Mechanical instability of adherens junctions overrides intrinsic quiescence of hair follicle stem cells. JF - Dev Cell Y1 - 2021 A1 - Biswas, Ritusree A1 - Banerjee, Avinanda A1 - Lembo, Sergio A1 - Zhao, Zhihai A1 - Lakshmanan, Vairavan A1 - Lim, Ryan A1 - Le, Shimin A1 - Nakasaki, Manando A1 - Kutyavin, Vassily A1 - Wright, Graham A1 - Palakodeti, Dasaradhi A1 - Ross, Robert S A1 - Jamora, Colin A1 - Vasioukhin, Valeri A1 - Jie, Yan A1 - Raghavan, Srikala AB -

Vinculin, a mechanotransducer associated with both adherens junctions (AJs) and focal adhesions (FAs), plays a central role in force transmission through cell-cell and cell-substratum contacts. We generated the conditional knockout (cKO) of vinculin in murine skin that results in the loss of bulge stem cell (BuSC) quiescence and promotes continual cycling of the hair follicles. Surprisingly, we find that the AJs in vinculin cKO cells are mechanically weak and impaired in force generation despite increased junctional expression of E-cadherin and α-catenin. Mechanistically, we demonstrate that vinculin functions by keeping α-catenin in a stretched/open conformation, which in turn regulates the retention of YAP1, another potent mechanotransducer and regulator of cell proliferation, at the AJs. Altogether, our data provide mechanistic insights into the hitherto-unexplored regulatory link between the mechanical stability of cell junctions and contact-inhibition-mediated maintenance of BuSC quiescence.

VL - 56 IS - 6 ER - TY - JOUR T1 - Scanning electron microscopy of murine skin ultrathin sections and cultured keratinocytes. JF - STAR Protoc Y1 - 2021 A1 - Banerjee, Avinanda A1 - Biswas, Ritusree A1 - Lim, Ryan A1 - Pasolli, Hilda Amalia A1 - Raghavan, Srikala AB -

Generating high-quality electron microscopy images of the skin and keratinocytes can be challenging. Here we describe a simple protocol for scanning electron microscopy (SEM) of murine skin. The protocol enables characterization of the ultrastructure of the epidermis, dermis, hair follicles, basement membrane, and cell-cell junctions. We detail the specific steps for sample preparation and highlight the critical need for proper orientation of the sample for ultrathin sectioning. We also describe the isolation and preparation of primary keratinocyte monolayers for SEM. For complete details on the use and execution of this protocol, please refer to Biswas et al. (2021).

VL - 2 IS - 3 ER -