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. 2018 Oct 29;19(12):e46433. doi: 10.15252/embr.201846433

Figure EV3. Characterization of pSer335 XErp1 antibody.

Figure EV3

  1. CSF extract was treated with Myc‐XErp1 IVT carrying the indicated combinations of the mutations DSG− (S33N S38N), DSA− (S284N S288N), ZBR− (C583A) and CaMKII− (T195A). Calcium was added, samples were taken at the indicated time points and as indicated treated with λ‐phosphatase. Samples were immunoblotted for the Myc‐tag and cyclin B2. The cyclin B2 membrane was stripped and reprobed for α‐tubulin. Asterisk indicates unspecific bands. Several lanes were removed at the dashed line.
  2. CSF extract was treated with Myc‐XErp1 CaMKII− ZBR− (T195A C583A) IVT at the indicated dilutions. An empty IVT not expressing XErp1 and an untreated condition were used as controls. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, pSer335 XErp1, and α‐tubulin. The XErp1 membrane was stripped and reprobed for the Myc‐tag. Asterisks indicate unspecific bands.
  3. CSF extract was treated with Myc‐XErp1 CaMKII− ZBR− (T195A C583A) IVT that was either wild‐type or mutated to alanine at Ser335. An empty IVT reaction not expressing Myc‐XErp1 served as control. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, the Myc‐tag, pSer335 XErp1, and α‐tubulin. Asterisks indicate unspecific bands.

Source data are available online for this figure.