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. 2016 Aug 19;7:12429. doi: 10.1038/ncomms12429

Figure 6. rRNA maturation defects and nucleolar stress in circANRIL-overexpressing cells.

Figure 6

(a) Relative quantification of pre-rRNA in circANRIL- or in circHPRT1-overexpressing HEK-293 cells using isoform-specific qPCRs (RNA from a pool of three biological replicates, quadruplicate measurements). (b) Pre-rRNA levels after RNAi against circANRIL or using scrambled siRNA control (quadruplicate measurements per condition). (c) Immunofluorescent staining of PES1 in circANRIL- or in circHPRT1-overexpressing HEK-293 or empty vector control cells. Quantification of (d) nucleoli and (e) nucleolar size in circANRIL- or circHPRT1-overexpressing or empty vector control cells (*** P<0.001). Data are given as mean±s.e.m. (analysis of variance, and Tukey as post hoc test). (f) Immunofluorescent staining of p53 (red) in circANRIL- or circHPRT1-overexpressing or empty vector control cells. (g) Schematic of transcriptome and proteome analyses in circANRIL-expressing HEK-293 cells and in control cells by genome-wide expression arrays and by pulseSILAC, respectively. For results of pathway analyses and procedure, see Supplementary Tables 4–6 and Methods section, respectively. L/M/H, light/medium/heavy medium.