Figure 2. (PR)₂₀ peptides impair reactions using RNA substrates in vitro and in vivo .

• A
  Reverse transcription of RNA (500 ng) with oligo dT in the presence of increasing doses of (PR)₂₀. Data represent the fluorometric quantification of the resultant cDNA (n = 3), mean values ± SD.
• B
  Percentage of RNA (1 μg) remaining after a 15′ digestion with increasing doses of RNase A in the presence or absence of (PR)₂₀ (5 μM; n = 3). Data represent mean values ± SEM.
• C, D
  High‐throughput microscopy (HTM)‐mediated analysis of 5‐ethynyl‐uridine (EU) (C) and polyA⁺ mRNA (D) levels per nucleus found in U2OS cells exposed to 10 μM (PR)₂₀ for 16 h. EU was added 30′ prior to fixation (n = 3). Black lines indicate median values in both panels. In (D), boxes define upper and lower quartiles and whiskers mark the highest and lowest observations.
• E
  Effect of (PR)₂₀ (10 μM, 16 h) in the number of Cajal Bodies (CB) identified by immunofluorescence with anti‐COILIN and anti‐SMN antibodies (n = 3). Data represent mean values ± SD.
• F
  Accumulation of Sindibis virus (SV) genomic RNA in BHK‐21 cells in the absence or presence of increasing doses of (PR)₂₀. The amount of viral and cellular RNA was quantified from total RNA isolated 4 h after infection by quantitative real‐time PCR (qRT–PCR) with specific primers (n = 3). Data represent mean values ± SEM.

Data information: **P < 0.01; ***P < 0.001; ****P < 0.0001; t‐test. Representative images from these analyses (C–E) are provided in each case. Scale bar (white) indicates 2–5 μm. See also Appendix Fig S1.