Journal: The American journal of pathology

Article Title: Elevated Expression of Moesin in Muscular Dystrophies.

doi: 10.1016/j.ajpath.2016.11.013

Figure Lengend Snippet: Figure 4 A: Radixin is synthesized by myofibroblasts. Diaphragms of mdx mice were double immunostained with radixin (red) and prolyl 4 hydroxylase b (P4Hb; green) antibodies. Note the colocalization of both proteins (P4Hb, green arrows; radixin, red arrows). B: Ezrin in mouse and human muscle biopsy specimens. Left panels: High resolution of myofibers from diaphragms of C57 (control) and mdx mouse and a low-resolution image showing a large area of mdx diaphragm. Right panels: Quadriceps femoris muscle of a healthy control and a 4-year-old DMD patient. Immunostained with ezrin antibodies (red), and nuclei were stained with DAPI (blue). Arrowheads point to myofibers’ central nuclei. Note that in the mdx mouse and in the DMD patient, ezrin colocalized with the myofiber membrane. Q17 Scale bars: 5 mm (top panel); 50 mm (bottom panel).

Article Snippet: Polyclonal moesin for double immunostaining (1:50), radixin (1:50), and monoclonal Pax7 (1:50) antibodies were from Proteintech (Rosemont, IL).

Techniques: Synthesized, Control, Staining, Membrane

Journal: Breast Cancer Research : BCR

Article Title: LIN7A is a major determinant of cell-polarity defects in breast carcinomas

doi: 10.1186/s13058-016-0680-x

Figure Lengend Snippet: Polarity abnormalities in invasive micropapillary carcinoma (IMPC). a Representative views of cell-division control protein 42 (CDC42), cis-Golgi marker (GM130) and occludin (OCLN) immunostainings in three normal ducts ( left panel ) and in three IMPC ( right panel ). Scale bars 10 μm, L lumen. G × 400 magnification. b Analysis of polarity protein expression and subcellular localization in 24 IMPC. Expression and cellular localization (sub-apical, apical, cytoplasmic, membranous, cell/cell: lateral membrane staining between two cells and basolateral) of polarity proteins was compared to that in normal cells. p-ERM phospho-ezrin-radixin-moesin, p-aPKCζ phospho-atypical PKC, PALS1 protein associated with Lin seven 1, SCRIB Scribble, ZO-1 zonula occludens 1

Article Snippet: Antibodies used were directed against phospho-ezrin-radixin-moesin (p-ERM,1:200 dilution, pH6.1, BD Biosciences), GM130 (clone 35, 1:100 dilution, pH9, BD Biosciences), cell division control protein 42 (CDC42, 1:300 dilution, pH6.1, Lifespan Biosciences, Seattle, WA, USA), phospho-atypical PKCζ (p-aPKCζ, clone190D10, 1:250 dilution, pH6.1, Cell Signaling), protein associated with Lin seven 1 (PALS1, 1:50 dilution, pH9, Lifespan Biosciences), occludin (OCLN, 1:200 dilution, pH6.1, Lifespan Biosciences), Scribble (SCRIB, 1:50 dilution, pH6.1, Santa Cruz Biotechnology, Dallas, TX, USA), zonula occludens 1 (ZO-1, 1:500 dilution, pH6.1, BD Biosciences), β-catenin (clone 14/Beta-catenin, 1:200 dilution, pH6.1, BD Biosciences), E-cadherin (clone 4A2C7, 1:100 dilution, pH9, Invitrogen, Camarillo, CA, USA), KI-67 (clone MIB1, 1:100 dilution, pH6.1, Dako A/S) or active-caspase-3 (1:250 dilution, pH6.1, Cell Signaling).

Techniques: Marker, Expressing, Staining

Journal: bioRxiv

Article Title: Radixin modulates stereocilia function and contributes to cochlear amplification

doi: 10.1101/2020.02.11.944355

Figure Lengend Snippet: . ( A ) Schematic diagram showing the putative function of radixin in stereocilia. ( B ) A low magnification image of the temporal bone preparation. Note the apical opening used for imaging. ( C ) Release of the dye di-3-ANEPPDHQ into the endolymphatic space stained Reissner’s membrane as well as the hair bundles. ( D ) Outer hair cell (OHC) stereocilia imaged during sound stimulation at 220 Hz, 80 dB SPL. ( E ) Representative confocal images of sections of the organ of Corti labelled with a radixin-specific monoclonal antibody (green) as well as phalloidin (red, staining actin), and overlay. The bundles of the sensory hair cells are intensely labeled by the radixin antibody. OHC 1, 2, 3 indicate the three rows of outer hair cells. Images were taken from the surface preparations of the apical turn. ( E ’) Inset showing a higher magnification view. ( F ) Three-dimensional reconstruction of the organ of Corti. A close-up on the inner hair cell area shows absence of radixin label in the cell bodies of the inner hair cells. Likewise, no radixin label was detected in the neuronal or synaptic region of the inner hair cells (right side). ( G ) A 3D reconstruction of outer hair cell stereocilia showing predominance of radixin labeling near the stereocilia base and consistent actin labeling in the hair cell body and stereocilia bundles. ( H ) Normalized average signal intensity profiles for radixin and actin expression (average of 11 bundles from 3 different animals) which shows decline in radixin labeling toward the tip of stereocilia and consistent actin labeling by phalloidin throughout the stereocilia. ( I ) Scatter plot showing lack of relation between radixin and phalloidin (staining actin) pixel intensities. A.u., arbitrary units.

Article Snippet: Permeabilization was followed with blocking step by incubating the samples for 2 hr in PBS containing 3% normal goat serum (NGS, 927503, BioLegend) and 3% BSA and then stained overnight at 4°C with the primary monoclonal antibody (mouse anti-radixin, ABNOH00005962-M06, Abnova) at a dilution of 1:500.

Techniques: Imaging, Staining, Membrane, Labeling, Expressing

Journal: Breast Cancer Research : BCR

Article Title: A novel mechanism of regulating breast cancer cell migration via palmitoylation-dependent alterations in the lipid raft affiliation of CD44

doi: 10.1186/bcr3614

Figure Lengend Snippet: Schematic representation of the proposed model of regulation of breast cancer cell migration via CD44 localisation in lipid rafts. When CD44 is affiliated with lipid rafts via palmitoylation of its cysteine residues, it is sequestered from binding its cytoplasmic binding partners and thus migration is restrained. However, when CD44 translocates outside lipid rafts in its de-palmitoylated state, its cytoplasmic tail is free to bind its cytoskeletal partners, subsequently facilitating cell migration. ERM, Ezrin/Radixin/Moesin.

Article Snippet: Antibodies against radixin, moesin, annexin II and merlin (rabbit) were from GeneTex, Inc. (Irvine, CA, USA).

Techniques: Migration, Binding Assay