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Fig. 4 | Acta Neuropathologica Communications

Fig. 4

From: Co-occurrence of chronic traumatic encephalopathy and prion disease

Fig. 4

Immunoblot study of PrPD brain regional characteristics. a-c: Immunoblots were carried out with proteinase K (PK)-treated brain homogenates (BH) obtained from the indicated brain regions sampled from the three study cases (CTE MV1-2C, CTE MV2K-C and CTE MM1) and sCJD controls (MV1, MV2C, MV2K and MM1). Different sample volume loads or exposures were used, to obtain a comparable representation of PK-resistant PrPD (resPrPD) bands. Membranes were probed with the indicated Abs to PrP: 3F4 to both resPrPD 1 and 2, 12B2 and 1E4 preferentially to type 1 and 2, respectively; SAF70 to the proximal C-terminal region. d: resPrPD loaded at higher concentrations (~ 1 mg tissue equivalent) in all lanes and were probed with SAF70. a and d: In CTE MV1-2C case 1, Ab 3F4 revealed resPrPD type 1 in all brain regions either as the sole component or associated with a higher mobility band in frontal (FC) and entorhinal cortices (EC) consistent with resPrPD type 2; resPrPD type 2 only was observed in the temporal cortex (TC). This type distribution was confirmed by probing with Ab 12B2 (to PrPD type 1) and by Ab 1E4 (preferentially to type 2). Ab SAF70, showed two fragments of 18.5 and 17 kDa commonly associated with resPrPD types 1 and 2, respectively, (solid arrowhead) and the C-terminal fragment CTF 13 kDa (asterisk). These fragments were visible, in the study case and controls BH at higher BH concentration (d). b and d: In CTE MV2K-C case 2, 3F4 showed the presence of a doublet of 19 kDa (dotted arrow) and 20 kDa (solid arrow) in all regions as typically seen in sCJDMV2K, with the exception of the cerebellum, which presented the 19 kDa band together with a fragment of about 18 kDa. Abs 12B2 and 1E4 detected the 20 kDa and 19 kDa component of the doublet, respectively. In putamen the 20 kDa fragment was detected only at the longer exposure (data not shown). SAF70 did not detect any additional fragments in the study case and sCJDMV2K control. c and d: In CTE case 3 MM1, 3F4 showed the presence of PrPD type 1 (21 kDa) in all different brain regions, which was confirmed with 12B2 while 1E4 showed no type 2 (blue arrow: non-specific band); SAF70 demonstrated the 18.5 kDa and CTF-13 fragments, typically present in sCJDMM1(MV)1. PC: parietal cortex (cx); OC: occipital cx; HI: hippocampus; CN: caudate nucleus; PUT: putamen; TH: thalamus; CE: cerebellum

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