Alexa fluor 594 conjugated goat anti rabbit igm

Liver tissues were fixed in 10% formaldehyde solution at 4 °C for paraffin sections or placed in 30% sucrose solution and embedded with liquid nitrogen-cooled isopentane for frozen sections. Paraffin sections of liver tissue were stained with hematoxylin and eosin (H&E) for light microscopy. For Oil Red O staining, frozen sections were incubated with the dye for 15 min and washed with 1× PBS. For immunofluorescence analysis, primary hepatocytes were stained overnight at 4 °C with antibodies against NCoR1 (Cell Signaling Technology) or PPARα (Santa Cruz Biotechnology). After washing with PBS, the sections were incubated with secondary antibodies (Alexa Fluor 488-conjugated goat anti-mouse IgG1 and Alexa Fluor 594-conjugated goat anti-rabbit IgM; Thermo Fisher Scientific) for 1 h at 37 °C and then counterstained with DAPI. NCoR1 and PPARα interactions were detected using a Duolink Proximity Ligation Assay kit (#DUO96010, Sigma-Aldrich).

Liver tissues were fixed in 10% formaldehyde solution at 4 °C for paraffin sections or placed in 30% sucrose solution and embedded with liquid nitrogen-cooled isopentane for frozen sections. Paraffin sections of liver tissue were stained with hematoxylin and eosin (H&E) for light microscopy. For Oil Red O staining, frozen sections were incubated with the dye for 15 min and washed with 1× PBS. For immunofluorescence analysis, primary hepatocytes were stained overnight at 4 °C with antibodies against NCoR1 (Cell Signaling Technology) or PPARα (Santa Cruz Biotechnology). After washing with PBS, the sections were incubated with secondary antibodies (Alexa Fluor 488-conjugated goat anti-mouse IgG1 and Alexa Fluor 594-conjugated goat anti-rabbit IgM; Thermo Fisher Scientific) for 1 h at 37 °C and then counterstained with DAPI. NCoR1 and PPARα interactions were detected using a Duolink Proximity Ligation Assay kit (#DUO96010, Sigma-Aldrich).

Paraffin sections of liver tissue (5 μm) were stained with haematoxylin and eosin (H&E) for light microscopy. Five random sections were investigated per slide to measure the necrotic area and determine the percentage of apoptotic cells. To measure the necrotic areas, sections were observed under an Axiovert 40 CFL microscope (Carl Zeiss, Oberkochen, Germany) and measured using iSolution DT 36 software (Carl Zeiss). For immunofluorescence staining, after deparaffinisation, sections were immunostained with antibodies against F4/80 (ab6640), Ly6G (ab25377), and 4-hydroxynonenal (4-HNE, ab46545) (all from Abcam). TUNEL staining was performed with commercial kits (Promega). After washing with phosphate buffered saline (PBS), secondary antibodies (Alexa Fluor 488-conjugated goat anti-mouse IgG1 and Alexa Fluor 594-conjugated goat anti-rabbit IgM, Thermo Fisher Scientific) were incubated for 1 h at 37 °C. Sections were counterstained with DAPI.