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. 2022 Mar 7;298(4):101770. doi: 10.1016/j.jbc.2022.101770

Figure 3.

Figure 3

Medium additives affecting the production of protease-resistant S3-PrP.A, S3 cells were grown in DMEM alone or in Opti-MEM with or without calcium (Ca)–EDTA. B, diminution of production of PK-resistant PrP following titrated addition of Ca–EDTA. Cell lysates were treated with 50 μg/ml PK. C, copper addition to S3-PrP cells propagated in Opti-MEM increased the yield of protease-resistant PrP. D, S3 cell extracts from cells grown in DMEM or Opti-MEM were digested (+) or not digested (−) with 50 μg/ml PK, and then samples were treated with PNGase F. The third lane represents a higher exposure of the second lane. E, S3 cells were grown in Opti-MEM, in DMEM alone, or in DMEM supplemented with 1.12 μM ZnCl2, 40 μM transferrin, 0.85 μM Cbl, or 40 μM lipoic acid. Cell lysates were treated with 50 μg/ml PK and PNGase F. Cbl, cobalamin; DMEM, Dulbecco's modified Eagle's medium; PK, proteinase K; PNGase F, peptide-N-glycosidase F; PrP, prion protein.