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At the time of incision (0 h), after 24 and 48 h of incubation with treatments, three photos per well were acquired under an optical microscope (Olympus IX51 model) at a magnification of 4×

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However, the simple linear interactions are unlikely and should be placed in the larger context of local regulatory networks including bidirectional communication with CRH/POMC/MSH&ACTH axis, HIF-1α, NF-κβ, MITF, PKA, PKC, second messengers, phosphorylation cascades, and nuclear receptor signaling (including vitamin D) that would act in a nonlinear fashion involving direct and indirect effects