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. Author manuscript; available in PMC: 2022 Feb 1.
Published in final edited form as: J Comp Neurol. 2020 Jun 27;529(3):481–500. doi: 10.1002/cne.24959

Figure 1. Antibody validation of anti-FMRP (PA8263) and anti-p-FMRP (P1125).

Figure 1.

a: Western blot assay on homogenized chick dorsocaudal brainstem samples. Molecular weight standards (left) were used to determine relative sizes of labeled proteins. Anti-FMRP (PA8263) detects a single band of just above of the 72 kDa at 1:1000 (left) and 1:4000 (right) antibody concentrations. b: Anti-FMRP (PA8263) detects the same band regardless of phosphatase treatment. Anti-p-FMRP (P1125) detects a band of the same molecular weight under normal conditions but is absent following phosphatase treatment. β-actin was used for loading control. c-d: Immunostaining of anti-FMRP (PA8263) following genetic knockdown of FMRP expression in NM neurons. Images were taken from a E19 chicken embryo that received in ovo electroporation of a Fmr1-shRNA with an EGFP reporter (X. Wang, Zorio, Schecterson, Lu, & Wang, 2018). Transfected NM neurons (green; dashed circles) show reduced FMRP immunostaining as compared to non-transfected neurons (magenta; solid circles). Scale bar = 20 μm in d (applies to c-d).