Research Paper Advance Articles

Figure 2. WRN regulates main NAD⁺ synthetic proteins and shares many transcription targets with NMNAT1-3. (A) Representative western blots of RIPA extracts from HEK293 parental cells with either 30 nM Scramble siRNA (Scr) or 30 nM WRN siRNA (WRN-KD) followed by 24 h 1 mM NR treatment. (B–F) Quantification of western blots from 4–6 biological repeats. Statistical analysis was performed in GraphPad Prism with either One-way ANOVA with Šidák multiple comparison’s test or Student’s t-test. (C) NMNAT1 was significantly decreased in WRN-KD cells compared to Scr (Veh) and Scr (NR) (One-way ANOVA, Tukey’s multiple comparisons test, p-value = 0.0108, student’s t-test p-value = 0.0163), (D) NAMPT was significantly increased in WRN-KD (Veh) cells compared to Scr (Veh) (One-way ANOVA, Tukey’s multiple comparisons test, p-value = 0.0492, Student’s t-test, p-value = 0.0010) and (E) NADSYN1 was upregulated in WRN-KD (Veh) compared to Scr (Veh) (Student’s t-test, p-value = 0.0442). (F–I) Venn diagram showing the shared transcription targets of WRN and NMNAT1-3 based on the ENCODE transcription database.