Based on in vitro evidence that 1a-OHase expression in macrophages is induced by TLR recognition of bacteria, we hypothesized that 1aOHase expression would be upregulated in both macrophages and mammary tissue during a mammary infection. Using an intramammary infection as a model in vivo bacterial infection, we present the first in vivo evidence that are at the site of a bacterial infection. The subsequent large increased expression of 24hydroxylase at the infection site supports local in vivo. Numerous in vitro studies have shown that 1a-OHase expression and subsequent 1,252D3 induction of 24-OHase in monocytes and macrophages is induced by TLR signaling in vitro. However, there was a lack of in vivo evidence that the vitamin D pathway was induced in macrophages in response to infection. Genes of the vitamin D pathway were elevated in macrophages in lesions of leprosy patients but that evidence could not confirm whether or not the pathway was upregulated in response to infection. In this study, we give in vivo confirmation that genes of the vitamin D signaling pathway were upregulated in response to bacterial infection. Biomimetic membrane systems have been developed to study, in controlled conditions, the biological events occurring at the cell membrane interface. Over the past 25 years, biomimetic models have been continuously improved with the aim of better mimicking the natural environment of biological membranes while allowing deeper investigations of 10-Gingerol membrane processes with various surface sensitive techniques such as Surface Plasmon Resonance, Atomic Force Microscopy, Quartz Crystal Microbalance, neutron-reflectometry, etc… Introduction of tethered supported bilayers has been a major implementation toward the reconstitution of integral membrane proteins within a fluid hydrophobic environment that preserves their functional properties. These supported lipid bilayers have been widely used to characterize interaction of ligands with membranes,6-gingerol dynamics of membrane proteins or even more complex receptor/ligand mediated intercellular contacts. Yet, in most cases, these reconstituted assemblies involved only the extracellular domains of cell receptors that were attached to the membrane bilayer in a manner preserving their lateral mobility, but did not take into account the underlying cytosolic compartment. Here, we present an improved model of biomimetic tBLM mimicking the three-dimensional architecture of a genuine biological membrane in that it defines a physical boundary between two distinct compartments, i.e. cis and trans sides. This was achieved by assembling a continuous tethered bilayer over a surface derivatized with the protein calmodulin to serve as a specific cytoplasmic marker. CaM is a ubiquitous, highly conserved intracellular Ca2+ sensor, capable of binding and regulating diverse intracellular targets such as protein kinases, protein phosphatases, phosphodiesterases, and ion channels. We established and validated the experimental conditions to assemble such a multilayered structure that preserves the functional activity of CaM and ensures the formation of a continuous yet fluid lipid bilayer acting as a proteinimpermeable barrier between two distinct compartments. The simple and robust procedure elaborated here to assemble multilayered biomimetic structures will be instrumental to reconstitute multimolecular complexes involving both cytosolic and membrane embedded proteins such as those implicated in many cell signaling pathways and will also be useful to characterize protein translocation across membranes.