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. 2001 Apr;183(7):2241–2248. doi: 10.1128/JB.183.7.2241-2248.2001

FIG. 3.

FIG. 3

Extraction of cryptococcal ribosomes with KCl and LiCl. (A) Dot blots of ribosomes from S. cerevisiae and C. neoformans. Lane 1, 0.6 μg of recombinant CnEF-2; lane 2, 0.5 μg of unwashed S. cerevisiae ribosomes; lane 3, 0.5 μg of S. cerevisiae ribosomes washed with 0.5 M KCl; lane 4, 0.5 μg of unwashed ribosomes from C. neoformans; lanes 5 to 7, 0.5 μg of C. neoformans ribosomes washed with 0.5 M KCl (lane 5), 1 M LiCl (lane 6), or 2 M LiCl (lane 7). Blots were treated with anti-CnEF3 antibody (1:1,000) and HRP-conjugated anti-mouse Ig as described in Materials and Methods. (B): Cryptococcal ribosomes were washed three times with 0.5 M KCl and subjected to sucrose density centrifugation as described in Material and Methods. Fractions were assayed for protein and CnEF3 protein by ELISA. 40S and 60S refer to expected positions of cryptococcal ribosomes based on centrifugation of purified S. cerevisiae ribosome subunits.