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Hypothyroidism In hypothyroidism, MTHFR mutations can impair the methylation processes required for converting T4 to T3, compounding the symptoms of fatigue, depression, and weight gain

4060 g of protein per sample was electrophoresed and transferred onto nitrocellulose membrane (Bio-Rad) for immunoblotting using antibodies against IRE1 (3294, Cell Signaling), XBP1 (ab220783, Abcam), PERK (3192, Cell Signaling), phospho-GCN2 (ab75836, Abcam), GCN2 (3302, Cell Signaling), phospho-eIF2 (9721, Cell Signaling), eIF2 (2103, Cell Signaling), ATF4 (11815, Cell Signaling), ATF6 (CosmoBio, BAM-73-500-EX), GCLC (sc-390811, Santa Cruz Biotechnology), GCLM (sc-22754, Santa Cruz Biotechnology), GPX4 (MAB5457, R&D Systems), SLC7A11 (12691, Cell Signaling), ACSL4 (SAB2701949, Sigma Aldrich), phosphor-JNK (9251, Cell Signaling), JNK (9252, Cell Signaling), LC3B (NB600-1384, Novus), Caspase-3 (9662, Cell Signaling), luciferase (NB600-307, Novus Biologicals), calreticulin (ab2907, Abcam), HSP90 (sc-13119, Santa Cruz Biotechnology), GAPDH (8884, Cell Signaling) and -actin (sc-1615, Santa Cruz Biotechnology) all with 1:1000 dilution in 3% (w/v) BSA (Sigma Aldrich)