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USP46 binds luxY. 14 / 14
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"To generate YFP-USP12 delMEIL , YFP-USP46 +MEIL and YFP-WDR20 deletion mutants, the corresponding DNA sequences were amplified by PCR and cloned into pEYFP-C1 using BamHI/HindIII."
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"A statistically significant, but very limited, increase in the N/C ratio of YFP-USP46 was noted."
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"The most striking finding was the relocation of YFP-USP12, but not YFP-USP46 to the PM when co-expressed with Myc-WDR20."
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"In line with previous studies ( Urbé et al., 2012 ; Lehoux et al., 2014 ), we found that YFP-USP12 and YFP-USP46 were located predominantly in the cytoplasm of 293 T and HeLa cells when expressed alon[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"YFP-USP46 plasmid was obtained by subcloning USP46 cDNA from a previously described plasmid ( Olazabal-Herrero et al., 2015 ) into pEYFP-C1 using BamHI/HindIII restriction sites."
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"We used a similar knockdown/overexpression approach to asses a potential effect of WDR20 on the localization of YFP-USP12 and YFP-USP46."
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"Image analysis showed that the nuclear to cytoplasmic ratio of YFP-USP46 was significantly higher when expressed with Myc-WDR20 NESm than with wild type Myc-WDR20 (Additional File 4b), suggesting that[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"Similar to UAF1 knockdown, WDR20 siRNA had no obvious effect on the localization of YFP-USP12 or YFP-USP46 ( Fig. 2 a)."
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"Whereas YFP-USP46 and Myc-WDR20 co-localized diffusely throughout the cytoplasm, YFP-USP12 and Myc-WDR20 co-localized at the cell periphery ( Fig. 2 b)."
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"In order to asses a potential effect of UAF1 on the localization of YFP-USP12 and YFP-USP46, cellular levels of UAF1 were either reduced using small interfering RNA (siRNA)-mediated knockdown or incre[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"As shown in Fig. 1 b, UAF1 knockdown had no obvious effect on the localization of YFP-USP12 or YFP-USP46."
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"On the other hand, since YFP-USP46 was not recruited to the PM, but accumulated in the cytoplasm when co-expressed with Myc-WDR20 (see Fig. 2 b), we tested the role of WDR20 NES on the localization of[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"However, YFP-USP46 +MEIL remained in the cytoplasm when co-expressed with Myc-WDR20 ( Fig. 3 e)."
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"On the other hand, YFP-USP12 and YFP-USP46 co-localized with UAF1-mRFP throughout the cytoplasm in co-transfection experiments ( Fig. 1 c) and, as shown by image analysis, co-expression with UAF1 mark[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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