PMC

Ribosomal sedimentation profiles of HEK-293 and HTB-14 cells. (Top panels) A₂₅₄ absorption spectra of 10 to 50% sucrose gradients containing lysates generated from mock-infected HEK-293 cells (A), lysates generated from HTB-14 cells (F), or HEK-293 lysates treated with EDTA (D). (Middle panels) A₂₅₄ absorption spectra of lysates generated from PV-RIPO-infected HEK-293 (B), PV-RIPO-infected HTB-14 (G), or PV-infected HEK-293 (E) cells. (C) A₂₅₄ absorption spectra of a 5 to 20% sucrose gradient containing free protein and 40S ribosomal subunit components from a PV-RIPO-infected HEK-293 cell lysate. Lysates from infected cells were generated at 4 hpi. Western blot analyses were performed with α-DRBP76, α-NF45, α-RpS6, and α-tubulin antibodies, as indicated. Viral cDNA corresponding to the IRES region was amplified by RT-PCR, and rRNA was analyzed from total RNA. M, monosome; P, polysome.

Neither Jjj1 nor Rei1 is required for tethering of its binding partner to the ribosome. Δrei1 and Δjjj1 cells, grown at 30 °C, were lysed in Buffer I. Ten A₂₅₄ units were centrifuged through 5–50% sucrose gradients. Upper, the A₂₅₄ was monitored and plotted versus the time course of fraction collection. The positions of 40 S, 60 S, 80 S, and polysomes are labeled. Lower, fractions were tested by immunoblot analysis for the presence of Jjj1 in fractions from Δrei1 lysates (left) or of HA-tagged Rei1 in Δjjj1 lysates (right). Antibodies specific for Jjj1 and HA, respectively, were used.

HPLC elution profile (A₂₅₄) and radioactivity (disintegrations per minute) of the ethyl acetate-soluble metabolites formed by C. elegans in 4 days from 6-nitro[³H]chrysene. Fractions eluting from the column were collected at 0.5-min intervals, and the radioactivity was measured by liquid scintillation counting. The inset shows the disappearance of 6-nitro[³H]chrysene and formation of ³H-labeled metabolites in cultures of C. elegans.

Effect of turbidity of, and absorbance of UV by, buffer on the apparent UV resistance of B. anthracis Sterne spores. Spores were prepared and irradiated as described previously (7) and in the text. Survival curves were plotted based on dosimetry calculated by using UVCalc with no correction (circles) or with correction for the A₂₅₄ of the buffer used (triangles) or for the A₂₅₄ values of both the buffer and the cell suspension (squares). For comparison, a heavy dashed line represents the UV inactivation curve of spores of B. subtilis ATCC 6633, as determined by Hoyer (5).