Figure EV5. Rpb9 is conserved in eukaryotes.
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AProtein alignment of Rpb9 orthologs from Saccharomyces cerevisiae, Homo sapiens, Mus musculus, Drosophila melanogaster, and Caenorhabditis elegans.
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BHEK293T cells were transfected with siRNA targeting POLR2I for 72 h and then the mRNA levels of POLR2I were analyzed. Bars represent mean, error bars represent standard deviation, significance was determined by one‐way ANOVA (unpaired) followed by Tukey's multiple comparison test, ***indicates P < 0.001 (n = 5 biological replicates).
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CPOLR2I knockdown increased the protein levels of autophagy receptor p62 which was subject to autophagic degradation. HEK293T cells were transfected with siRNA targeting POLR2I for 72 h and then the protein levels of p62 were analyzed.
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DPOLR2I knockdown reduced the number of autophagosomes shown by LC3 puncta. HEK293T cells with expression of GFP‐LC3B were transfected with siRNA targeting POLR2I for 72 h and then the LC3 puncta was observed and quantified. Bars represent mean, error bars represent standard deviation, significance was determined by one‐way ANOVA (unpaired) followed by Tukey's multiple comparison test, **indicates P < 0.01 (n = 5 biological replicates). Scale bars: 10 μm.