Hypothalamic ER Stress Signaling Proteins

A subset of 6 mice per group were used for assessment of hypothalamic PERK, pPERK^Thr980, eIF2α, peIF2α^Ser55, IRE1α, pIRE1α^Ser724, ATF6, TNFα and tubulin protein expression using standard methodology, as previously described (McGavigan et al., 2015 (link)). Antibodies against PERK, pPERK^Thr980 and peIF2α^Ser55 were from Cell Signaling (3179 and 3398, respectively). Antibodies against eIF2α, ATF6 and tubulin were from Santa Cruz (sc-133227, sc-22799, sc-23948, respectively) and TNFα, IRE1α and pIRE1α^Ser724 antibodies were from Abcam (ab66579, ab37073, ab104157, respectively). All antibodies were used at a concentration of 1:1000 and pixel densities of immunoreactive bands were quantified using FluorChem 9900 (Alpha Innotech, CA).