Fig 1: Quantitative analysis shows elevated sPTPRZ in the CSF of glioma patients. (A and B) For quantification of cleaved PTPRZ, semiquantitative western blot analysis was performed using CSF samples from glioma (n = 24), MS (n = 27), schwannoma (SW, n = 14), and control (n = 21) patients. TTR levels in CSF samples were determined by ELISA. Data show the ratio of signal intensities of sPTPRZ-Long (A) and sPTPRZ-Short (B) compared with the levels of the internal standard protein TTR. Data are presented as the mean ± SEM, with individual data points represented by circles. N.S., not significant, *P < .05, ****P < .0001 by Kruskal–Wallis test with Dunn’s test. (C) As described for sPTPRZ-Long in (A) except the CSF samples were from patients with grade 1 (n = 3), 2 (n = 2), 3 (n = 12), and 4 (n = 7) glioma. N.S., Not significant by Kruskal–Wallis test with Dunn’s test. (D) Receiver operating characteristic curve analysis of the value of the sPTPRZ-Long to TTR signal intensity ratio in CSF for discriminating between glioma and MS patients. The area under the curve was 0.9676 (P < .0001).
Fig 2: Western blot analysis of PTPRZ and sPTPRZ in microsomal fractions and CSF from glioma patients reveals 2 forms. (A) CSF samples from glioma patients A and B were treated with or without chondroitinase ABC (Chase ABC) and probed with Santa Cruz or Sigma anti-PTPRZ antibodies. (B) Schematic of the structures of PTPRZ-Long, PTPRZ-Short, and their proteolytic cleaved forms, sPTPRZ-Long and sPTPRZ-Short, respectively. Regions recognized by the anti-PTPRZ1 (Sigma), anti-PTPRZ (Santa Cruz), and Cat-315 antibodies used are indicated. (C) Microsomal fractions prepared from glioma tissues and CSF samples were treated with or without chondroitinase ABC (Chase ABC) and used for western blot analysis with anti-PTPRZ (Santa Cruz), Cat-315, and anti-Aggrecan antibodies. Cadherin and TTR were probed as loading controls for microsomal fractions and CSF, respectively. A and B are from different patients. (D) Western blot analysis of CSF samples from a glioma patient treated with chondroitinase ABC (Chase ABC) or endo-ß-galactosidase and probed with anti-PTPRZ (Santa Cruz).
Fig 3: Western blot analysis shows elevated levels of sPTPRZ in the CSF of glioma patients. (A) Western blot analysis of CSF samples from patients with iNPH, glioma (glioblastoma [GBM] and anaplastic oligodendroglioma [AO]), multiple sclerosis (MS), or schwannoma (SW). Blots were probed with anti-PTPRZ (Santa Cruz; upper) or with Cat-315 antibody (lower). Transthyretin (TTR) was probed as a loading control. (B) Western blot analysis of CSF samples from control subjects (C1, facial spasm; C2, unruptured cerebral aneurysm; and C3, iNPH) and from glioma patients collected from cerebral cisterns before surgery (G1) and from ventricles after surgery (G1*) for sPTPRZ and TTR. Relative signal intensities of sPTPRZ and sPTPRZ/TTR are shown.
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