Immunohistochemical Analysis of Molecular Markers

Immunohistochemical analysis was performed as already described [41 (link),42 (link)]. Sections were incubated overnight with the following: anti-PPAR-α (1:1000, Santa Cruz Biotechnology, Heidelberg, Germany, sc-398394), anti-TRL4 (1:1000, Santa Cruz Biotechnology, Heidelberg, Germany, sc-293072), anti-claudin-11 (1:100, Thermofisher, Oxford, UK), or anti-occludin (1:100; Thermofisher, Oxford, UK). Images were collected using a Leica DM6 (Milan, Italy) microscope. The percentage area of immunoreactivity (described by the number of positive pixels) was reported as a percentage of the total tissue area [43 (link)].

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Antonuccio P., Marini H.R., Micali A., Romeo C., Granese R., Retto A., Martino A., Benvenga S., Cuzzocrea S., Impellizzeri D., Di Paola R., Fusco R., Cervellione R.M, & Minutoli L. (2021). The Nutraceutical N-Palmitoylethanolamide (PEA) Reveals Widespread Molecular Effects Unmasking New Therapeutic Targets in Murine Varicocele. Nutrients, 13(3), 734.

Publication 2021

anti-PPAR-α anti-TRL4 anti-claudin-11 anti-occludin

Claudin Microscope Occludin Ppar Tissue

Corresponding Organization : University of Messina

Other organizations : Royal Manchester Children's Hospital

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Variable analysis

independent variables

Anti-PPAR-α (1:1000, Santa Cruz Biotechnology, Heidelberg, Germany, sc-398394)
Anti-TRL4 (1:1000, Santa Cruz Biotechnology, Heidelberg, Germany, sc-293072)
Anti-claudin-11 (1:100, Thermofisher, Oxford, UK)
Anti-occludin (1:100; Thermofisher, Oxford, UK)

dependent variables

Percentage area of immunoreactivity (described by the number of positive pixels) as a percentage of the total tissue area

control variables

Sections were incubated overnight with the specified antibodies

controls

No positive or negative controls were explicitly mentioned in the provided information.

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