Sc 372 g

Protein extracts from cells or liver tissue were resolved by SDS-PAGE,
transferred onto nitrocellulose membranes, and incubated with antibodies
specific for human and mouse (human/mouse) p65 (1:1000, SC-372-G, Santa Cruz
Biotechnology or ab7970, Abcam), human/mouse pSer⁵³⁶-p65 (1:1000,
#3033, Cell Signaling), human/mouse pSer⁴⁶⁸-p65 (1:1000,
#3039, Cell Signaling), human/mouse pSer²⁷⁶-p65 (1:1000,
#3037, Cell Signaling,), human/mouse IκBα (1:1000,
#4814, Cell Signaling,), pERK (1:1000, #9106, Cell Signaling),
pp38 (1:1000, #9215, Cell Signaling), p38 (1:1000, #9212, Cell
Signaling), or pJNK (1:1000, #9251, Santa Cruz Biotechnology) before
being stripped and incubated with antibodies against actin (1:5000,
#691002, MP Biomedicals) or GAPDH (1:20000, G9295, Sigma-Aldrich).

Immunohistochemical staining was performed on paraffin liver sections
after 10% formalin fixation with primary antibody against p65 (1:200,
SC-372-G, Santa Cruz) and Alexa Fluor 594–conjugated secondary antibody
(Life Technologies). Nuclei were stained with Hoechst 33342 (Sigma Chemical
Co.). For immunocytofluorescence, primary MEFs were fixed with 4%
paraformaldehyde and stained with primary antibody against p65 (1:100, SC-372-G)
and Alexa Fluor 594–conjugated secondary antibody. Counterstaining was
performed Alexa Fluor 488–conjugated Phalloidin (Molecular Probes).
Nuclei were stained with Hoechst 33342.