Figure 5.

BV-associated bacterial supernatants affect vaginal epithelial barrier function in vitro and affect pathways involved in mTOR activity and epithelial function

(A) Heatmap showing differentially abundant proteins in VK2 cells treated with different bacterial culture supernatants for 24 h, as observed by mass spectrometry. Proteins that activate mTOR cluster in the group that is upregulated in BV-associated bacterial treated cells (red box). Numerous structural and signaling adherens proteins were also differentially regulated by BV-associated bacteria (highlighted in green).

(B) Pathway analysis showing top pathways including wound healing, tissue remodeling, and mTOR activation.

(C) G. vaginalis inhibited VK2 cell differentiation and thickness compared with L. crispatus (n = 9).

(D) G. vaginalis inhibited wound-healing ability as measured by scratch assay in Hec1a cells (n = 9).

(E) M. mulieris decreased barrier integrity as measured by electrical resistance (TEER) on transwell membranes of Hec1a cells (n = 12).

(F) M. mulieris induced a leaky barrier with protein-sized particles (dextran-FITC, 70 kDa) able to translocate across the membrane, but not larger-sized particles (n = 6).

All error bars represent standard deviation from the mean, p values represent unpaired t test results. See also Figure S4.