Skip to main content
. 2023 Jun 24;299(8):104951. doi: 10.1016/j.jbc.2023.104951

Figure 2.

Figure 2

Depletion of PAF49 results in the inhibition of rDNA transcription and cell proliferation.A, after cells were treated with 1 mM IAA for 3 h, they were pulsed with 5-ethynyl uridine (EU) for 15 min, and de novo synthesized RNA (red) was visualized as described against a DAPI background (94). To rescue rDNA transcription, wild-type mouse PAF49 (WT mPAF49) was ectopically expressed for 24 h before the cells were treated with IAA. Scale bar = 50 μm. B, cells containing AID-PAF49 were treated with 1 mM IAA or vehicle for 4 days. On days 0, 2, 3, and 4, live cells were counted as described (79). The growth curves were constructed with a nonlinear fit with GraphPad Prism. Significance between day 4 control and IAA data points was determined by a two-tail t test, p = 0.0003. C, trypan blue exclusion was used to count live cells and measure percentage viability within the cell population. A two-tail t test was performed to test for significance, p = 0.9762. D, after cells were treated with 1 mM IAA for 3 days, they were pulsed with ethynyl-2′-deoxyuridine (EdU) for 1 h, and de novo synthesized DNA (red) was visualized as described against a DAPI background (94). The ectopic expression of WT PAF49 rescued cell proliferation. WT mPAF49 was ectopically expressed for 24 h before the cells were treated with IAA. Scale bar = 100 μm.