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. 2018 Sep 11;37(19):e99023. doi: 10.15252/embj.201899023

Figure EV1. Validation of rabbit polyclonal α‐SerCT and α‐SerCT2 antibodies.

Figure EV1

  1. Amino acid sequence of predicted polySer RAN protein with the unique C terminus. Peptide sequences used to generate rabbit polyclonal antibodies are underlined.
  2. Schematic diagram of FLAG‐SerCT construct expressing an ATG‐initiated N‐terminal FLAG‐tagged polySer expansion protein followed by its endogenous C‐terminal sequence. Co‐localization of immunofluorescence (IF) staining using α‐FLAG (red) and α‐SerCT and α‐SerCT2 (green) in HEK293T cells transfected with FLAG‐SerCT but not preimmune serum.
  3. Immunoblots showing detection of recombinant polySer protein using α‐FLAG (left) and α‐SerCT (right) in the lysates of HEK293T cells transfected with FLAG‐SerCT (second lanes) but not pcDNA3.1 (first lanes).
  4. Immunochemistry of SCA8 mouse brain using α‐SerCT and α‐SerCT2 (left panels) antibodies shows similar punctate aggregates. Aggregates are not detected with respective preimmune sera (right panels).
  5. Immunochemistry using both α‐SerCT and α‐SerCT2 detect similar aggregates in SCA8 human autopsy tissue but not control cerebellum.

Source data are available online for this figure.