Fig. 5 | Cell Death & Differentiation

Fig. 5

From: UBAP2L arginine methylation by PRMT1 modulates stress granule assembly

Fig. 5

PRMT1 asymmetrically dimethylates UBAP2L by targeting the RGG motif. a Coomassie blue-stained gel showing that the Co-IP/MS assay identified the association of PRMT1 with UBAP2L. b Confirmation of the interaction between UBAP2L and PRMT1 by IP/WB. HEK293 cells were simultaneously transfected with Flag tag or Flag-UBAP2L plasmid, followed by WB analysis of endogenous PRMT1 in the Flag precipitates. c In vitro methylation assay on recombinant UBAP2L-mediated by PRMT1 via monitoring the ADMA level. Asterisk indicates a positive ADMA signal. The lower panel showed loading controls for recombinant UBAP2L and immunoprecipitated PRMT1. d Co-IP/WB analysis of the associations of the UBAP2L deletants with PRMT1. HEK293 cells were transfected with Flag-tagged UBAP2L-WT or indicated deletants, followed by treatment with/without AS (500 μM, 1 h), in combination with/without RNase A in the lysis buffer. e Co-IP/WB analysis of the associations of the UBAP2L mutants with PRMT1 and ADMA levels. HEK293 cells were transfected with Flag-tagged UBAP2L-WT or indicated mutants (depicted in Supplementary Fig. S1), followed by treatment with/without AS (500 μM, 1 h). The Flag precipitates were analyzed by WB for PRMT1 and ADMA. f SG competence in UBAP2L-KD HeLa cells exposed to the same treatments as in (e), using Flag (red)/eIF4G (green) as SG markers. g Quantification of SGs in (f). h SG competence in UBAP2L-KD HeLa cells with Flag-tagged UBAP2L-WT or the indicated point mutants (depicted in Supplementary Fig. S1). i Quantification of SGs in (h). j Co-IP/WB analysis of the associations of the UBAP2L point mutants with PRMT1 and ADMA levels in UBAP2L-KD HEK293 cells. n = 3. Scale bars = 10 μm

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