Immunohistochemical Profiling of Tissue Samples

Immunohistochemical analysis was performed as already described [49 (link)]. The sections were incubated overnight with primary antibodies: anti-TGF-β antibody (1:250, Santa Cruz Biotechnology), anti-(α-sma antibody (1:250, Santa Cruz Biotechnology), anti-nitrotyrosine antibody (1:500, Millipore), anti-PARP antibody (1:250, Santa Cruz Biotechnology), anti-CD4 (1:250, Santa Cruz Biotechnology) and anti-CD8 (1:250, Santa Cruz Biotecnology). All sections were washed with PBS and then treated as previously reported [50 (link)]. Stained sections from each mouse were scored in a blinded fashion and observed using a Leica DM6 microscope (Leica Microsystems SpA) following a typical procedure [51 (link)]. The histogram profile is related to the positive pixel intensity value obtained [52 (link)]. The number of positive cells was counted in three sections per animal and presented as the number of positive cells per high-power field.

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Fusco R., Siracusa R., D’Amico R., Cordaro M., Genovese T., Gugliandolo E., Peritore A.F., Crupi R., Di Paola R., Cuzzocrea S, & Impellizzeri D. (2020). Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Inhibitor as a Novel Therapeutic Tool for Lung Injury. International Journal of Molecular Sciences, 21(20), 7761.

Publication 2020

anti-TGF-β antibody anti-(α-sma antibody anti-nitrotyrosine antibody anti-PARP antibody anti-CD4 Leica DM6 microscope

Animal Anti antibody Antibodies Antibody Microscope Mouse Nitrotyrosine Tgf β

Corresponding Organization : Saint Louis University

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Variable analysis

independent variables

Primary antibodies used for immunohistochemical analysis: anti-TGF-β antibody, anti-α-sma antibody, anti-nitrotyrosine antibody, anti-PARP antibody, anti-CD4, anti-CD8

dependent variables

Positive pixel intensity value obtained
Number of positive cells per high-power field

control variables

Staining procedure as previously reported [50 (link)]
Scoring of stained sections in a blinded fashion
Observation using a Leica DM6 microscope following a typical procedure [51 (link)]

controls

Positive controls not explicitly mentioned
Negative controls not explicitly mentioned

Annotations

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