Skip to main content
. 2021 Oct 30;12(4):1913–1927. doi: 10.1016/j.apsb.2021.10.025

Figure 1.

Figure 1

PHF6 localizes to the nucleolus and regulates rDNA transcription. (A) Confocal images of HeLa cells transfected with GFP-PHF6 plasmids using nucleolin as a nucleolar marker. Scale bar, 10 μm. (B) Arrows indicate the position of qPCR primers on 47S pre-rRNA. (C) qPCR analysis of 47S pre-rRNA from HeLa cells transfected with Flag-empty vector (EV) or Flag-PHF6 plasmid. Western blots show the protein levels of Flag-PHF6. (D) Schematic representation of pHrD-IRES-Luc. (E) Flag-EV, Flag-PHF6, and PHrD-IRES-Luc were co-transfected into HeLa cells for 24 h, and the activity of luciferase was measured. (F) qPCR analysis of 47S pre-rRNA from HeLa cells silenced with negative control shRNA (shNC) and PHF6 shRNAs (shPHF6#1 and shPHF6#2). Immunoblot of PHF6 in the indicated HeLa cells. (G) Jurkat cells were silenced with negative shRNA (shNC), PHF6 shRNAs (shPHF6#1 and shPHF6#2), and PHrD-IRES-Luc was then transfected into the cells. Luciferase activity was measured after an additional 24 h. (H) Confocal images of Jurkat cells transfected with the GFP-PHF6 plasmid. Scale bar, 10 μm. (I) and (J) qPCR analysis of 47S pre-rRNA from Jurkat (I) and U937 cells (J) infected with negative shRNA (shNC), PHF6 shRNAs (shPHF6#1 and shPHF6#2), and over-PHF6. Immunoblot of PHF6 in the indicated Jurkat and U937 cells. GAPDH was used as loading control. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001. All the values are expressed as the mean ± SD, n = 3.