Skip to main content
. 2022 Dec 30;24(3):146–157. doi: 10.1111/tra.12877

FIGURE 6.

FIGURE 6

Depletion of Treacle reduces RABV production and RABV infection or P3 expression impairs rRNA synthesis. (A) HEK‐293T cells were transfected with scrambled (scr) or Treacle‐targeting (siTCOF1) siRNA 2 days prior to infection with RABV (MOI 0.01). RABV titers were measured at 24 and 48 h p.i. (mean TCID50/ml ± SEM, n = 3). Statistical analysis used Student's t‐test, ***, p = 0.0002. (B, C) HEK‐293 cells infected with RABV (MOI of 2) or (D, E) HeLa cells transfected to express RABV P3 or RABV P3‐KRm, were analysed to determine nucleolar EU incorporation (as described in the legend to Figure 3). EU reagent was added 1 h prior to fixation at 24 h p.t. or p.i. Infected cells (B, C) were detected by IF staining for RABV N protein, and transfected cells (D, E) by expression of GFP. Hoechst 33342 was used to detect nuclei and assist with identification of nucleoli. (C) Mean nucleolar EU fluorescence (arbitrary units [a.u.]) of mock‐ and virus‐infected samples (n ≥ 309 nucleoli for each condition from three independent samples). Microscope settings were identical for all samples. (E) Histogram of nucleolar EU fluorescence represented as relative to cells expressing GFP alone (control) (n ≥ 110 nucleoli for each condition, from three independent experiments); Statistical analysis used Student's t‐test; ***p < 0.001; ****p < 0.0001; ns, nonsignificant (p > 0.05).