Journal: Human mutation

Article Title: Biallelic loss of function variants in PPP1R21 cause a neurodevelopmental syndrome with impaired endocytic function.

doi: 10.1002/humu.23694

Figure Lengend Snippet: FIGURE 2 PPP1R21 co-localizes with the main early endosome protein EEA1 but not Golgi proteins and PPP1R21 staining is absent in fibroblasts obtained from patient 3_V:4 who is homozygous for c.1607dupT p.(Leu536Phefs*7). (A) In order to determine the entity of PPP1R21 positive vesicles, we performed co-localization studies using Golgi marker GM130 (mouse, ab169276, Abcam, USA) and 58K (ab27043 mouse, Abcam, USA) as well as EEA1 antibody to mark the early endosome. While no co-localization with Golgi markers was observed (upper panels), PPP1R21 (rabbit, HPA036792, Atlas antibodies, Sweden) nearly completely co-localized with EEA1 (lower panel). Scale bar 20 𝜇m. (B) Using two different PPP1R21 antibodies, HPA036792 (ab1, antibody epitope is represented by aminoacid aa161-256) and HPA 036791 (ab2, antibody epitope is represented by aminoacid aa572-666), we found complete loss of the vesicular staining pattern in PPP1R21 mutant fibroblasts we previously observed in control cells while no difference regarding EEA1 staining was detected between PPP1R21 mutant and control fibroblasts. Scale bar: 20 𝜇m

Article Snippet: See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense REHMAN ET AL. 277 F IGURE 2 PPP1R21 co-localizes with themain early endosome protein EEA1 but not Golgi proteins and PPP1R21 staining is absent in fibroblasts obtained from patient 3_V:4 who is homozygous for c.1607dupT p.(Leu536Phefs*7). (A) In order to determine the entity of PPP1R21 positive vesicles, we performed co-localization studies using Golgi marker GM130 (mouse, ab169276, Abcam, USA) and 58K (ab27043mouse, Abcam, USA) as well as EEA1 antibody tomark the early endosome.While no co-localization with Golgi markers was observed (upper panels), PPP1R21 (rabbit, HPA036792, Atlas antibodies, Sweden) nearly completely co-localized with EEA1 (lower panel).

Techniques: Staining, Marker, Mutagenesis, Control

Journal: Journal of Virology

Article Title: Anti-Glycoprotein H Antibody Impairs the Pathogenicity of Varicella-Zoster Virus in Skin Xenografts in the SCID Mouse Model

doi: 10.1128/jvi.01338-09

Figure Lengend Snippet: FIG. 8. Localization of gH and MAb 206 relative to EEA1 and Vps4 in fibroblasts in vitro. HELFs inoculated with pOka were mock treated (A to D, K to N) or antibody treated (F to I, P to S) for 48 h, fixed, permeabilized, stained, and examined by confocal microscopy. Uninfected HELFs were mock treated (E and O) or antibody treated (J and T) for 48 h, fixed, permeabilized, stained, and examined. Arrowheads highlight colocalization. (A) gH, red; EEA1, blue; TGN46, green; nuclei, gold. (B) MAb 206, red; EEA1, blue; TGN46, green; nuclei, gold. (C) gH, red; Vps4, blue; TGN46, green; nuclei, gold. (D) MAb 206, red; Vps4, blue; TGN46, green; nuclei, gold. Bar, 5 m.

Article Snippet: Cellular localization of VZV proteins was performed using primary antibodies to VZV proteins gH (SG3 monoclonal mouse anti-gH; Biodesign, Saco, ME), ORF23 (rabbit polyclonal) (7), and gE (rabbit polyclonal) (25) and to cellular proteins TGN46 (AHP500 polyclonal sheep anti-TGN46; AbD Serotec, Oxford, United Kingdom), early endosome antigen 1 (EEA1) (NB300-502 rabbit anti-EEA1; Novus Biologicals, Littleton, CO), and vacuolar protein sorting 4 (Vps4) (sc-32922 rabbit anti-Vps4; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: In Vitro, Staining, Confocal Microscopy