dsred rab11a wild type  (Addgene inc)

Journal: PLoS Biology

Article Title: Huntingtin Is Required for Epithelial Polarity through RAB11A-Mediated Apical Trafficking of PAR3-aPKC

doi: 10.1371/journal.pbio.1002142

Figure Lengend Snippet: (A) Mammary gland sections stained for E-cadherin and PAR3 or aPKC. Arrows indicate cytoplasmic accumulation and impaired localization of PAR3 and aPKC to the apical surface; asterisks indicate small lumens. (B) Mammary gland sections stained for keratin 5 (K5) and GM130. (C) Percentage of LCs showing ribbon-like and fragmented localization of GM130 (control: n = 4 mice; mutant: n = 4 mice). (D) Mammary gland sections stained for HTT 4C8 and PAR3 or aPKC. (E) HTT/PAR3/PAR6/aPKC/RAB11A complexes were immunoprecipitated from MCF-10A cells. Mouse IgG (mIgG) was used as a negative control. The immunoprecipitates (IP) were analyzed by western blotting. All scale bars, 10 μm. Error bars, SEM. *** p <0.001. Complete statistical analyses with number of measures are detailed in .

Article Snippet: GFP-RAB11A wild-type (WT), dominant-negative (S25N), and constitutively active (Q70L) (referred to herein as RAB11A WT , RAB11A S25N and RAB11A Q70L , respectively) were obtained from B. Goud (Institut Curie, France).

Techniques: Staining, Control, Mutagenesis, Immunoprecipitation, Negative Control, Western Blot

Journal: PLoS Biology

Article Title: Huntingtin Is Required for Epithelial Polarity through RAB11A-Mediated Apical Trafficking of PAR3-aPKC

doi: 10.1371/journal.pbio.1002142

Figure Lengend Snippet: (A) Mammary gland section stained for HTT 4C8 and RAB11A. (B) Mammary gland section stained for E-cadherin and RAB11A. (C) Twenty-four–hour and four-day MDCK 3-D cultures stained for HTT 4C8 and RAB11A. Colocalization of HTT and RAB11A is displayed in yellow (merge). (D) Twenty-four–hour and four-day MDCK 3-D cultures stained for RAB11A. HTTFL is tagged with mCherry and fluorescence is displayed in magenta and the colocalization of RAB11A and HTTFL appears in white. (E) Twenty-four–hour and four-day MDCK 3-D cultures transfected with RAB11A WT , RAB11A Q70L or RAB11A S22N , stained for PAR3. RAB11 is tagged with GFP and fluorescence is displayed in magenta. The colocalization of aPKC and RAB11A appears in white. (F) Representative line-scan analysis (relative fluorescence intensity; at least 20 cells were analyzed per condition). (G) Percentage of acini with normal lumen. (H) Quantification of acini size. (G and H) Control+RAB11A WT : n = 59 acini, Control+RAB11A Q70L : n = 54 acini, Control+RAB11A S22N : n = 66 acini, shHTT2+RAB11A WT : n = 60 acini, shHTT2+RAB11A Q70L : n = 72 acini, shHTT2+RAB11A S22N : n = 93 acini. (I) FM64-4 4-day MDCK 3-D structures were video recorded. Maximum intensity and z projections are shown. All scale bars, 10 μm. Error bars, SEM. ** p <0.01; *** p <0.001. Complete statistical analyses with number of measures are detailed in .

Article Snippet: GFP-RAB11A wild-type (WT), dominant-negative (S25N), and constitutively active (Q70L) (referred to herein as RAB11A WT , RAB11A S25N and RAB11A Q70L , respectively) were obtained from B. Goud (Institut Curie, France).

Techniques: Staining, Fluorescence, Transfection, Control

Journal: PLoS Biology

Article Title: Huntingtin Is Required for Epithelial Polarity through RAB11A-Mediated Apical Trafficking of PAR3-aPKC

doi: 10.1371/journal.pbio.1002142

Figure Lengend Snippet: During epithelial morphogenesis, HTT modulates the activation of RAB11A (1). HTT-RAB11A forms a complex with PAR3-aPKC, which may be recruited to HTT-kinesin 1 apical vesicles (2). HTT coordinates apical recycling of PAR3-aPKC vesicles (3). PAR3-aPKC accumulation at the pre-apical patches (PAP) (4) triggers the expansion of the apical membrane, leading to the formation of a central lumen (5).

Article Snippet: GFP-RAB11A wild-type (WT), dominant-negative (S25N), and constitutively active (Q70L) (referred to herein as RAB11A WT , RAB11A S25N and RAB11A Q70L , respectively) were obtained from B. Goud (Institut Curie, France).

Techniques: Activation Assay, Membrane