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Fig. 2 | Mobile DNA

Fig. 2

From: Properties of LINE-1 proteins and repeat element expression in the context of amyotrophic lateral sclerosis

Fig. 2

Nuclear localization of L1 ORF1 protein. a Endogenous ORF1p detected in 2102Ep cells by the α-4H1-ORF1 antibody. ORF1p is mostly cytoplasmic where it concentrates in granules and occasionally at the nuclear membrane. It is faintly seen in the nucleoplasm and concentrates in nucleoli of a subset of cells. b, c Exogenously expressed EGFP-tagged ORF1p strongly concentrates at the nuclear membrane and in perinucleolar foci of 5% or fewer human (b) U2OS or (c) HEK 293T cells with attendant reduction in size and number of cytoplasmic granules. Cotransfected mCherry-PSP1 marks nuclei and is excluded from nucleoli. d Endogenous ORF1p detected by α-4H1-ORF1 also forms discrete nuclear foci in a minor percentage of 2102Ep cells. Selected foci are enlarged in panels to the right. e Alu RNA, tagged with six MS2 coat protein recognition stem loops and expressed from construct pBS 7SL Alu-MS2 (Ya5), was detected by FISH using a Cy3-tagged DNA probe to the MS2 stem loops. Alu RNA colocalizes with nuclear foci marked by EGFP-tagged ORF1p in HEK 293T cells. f Nuclear foci of MS2 stem loop-tagged full-length SVA RNA detected by the Cy3-MS2 DNA probe do not colocalize with foci marked by ORF1p-EGFP. g RNA having 31 tandem G4C2 repeats detected by FISH using a Cy3-conjugated (C4G2)4 DNA probe induces intense intranuclear or cytoplasmic RNA aggregates that colocalize with ORF1p-EGFP in a minor percentage of HEK 293T cells (nuclear granules are marked by small arrows and cytoplasmic granules by large arrows). Size bars are 10 μm

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