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The membranes were blocked with 5% nonfat milk in TBST buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, and 0.1% Tween-20) for 2 h at room temperature, followed by incubation overnight at 4 C with the following primary antibodies: rabbit polyclonal anti-TEX44 (prepared in-house, 1:2000), mouse monoclonal anti-HA (MBL, m180-3, 1:5000), mouse monoclonal anti-FLAG (Sigma-Aldrich, F3165, 1:5000), rabbit polyclonal anti-CPT1B (abcam, ab134988, 1:2000), mouse monoclonal anti--Actin (Santa, sc-47778, 1:1000) or mouse monoclonal anti--Tubulin (ABclonal, AC021, 1:5000)

Step 2: Set Your Protein Target Aim for 1.6 to 2.0 grams of protein per kilogram of body weight per day (which is roughly 0.8 to 1 gram per pound)

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32 Furthermore, subsequent large retrospective studies have demonstrated no effect of GLP-1RA on the increased risk of ischemic optic neuropathy