Journal: Investigative Ophthalmology & Visual Science

Article Title: Transmembrane Mucin 1 Blocks Fluorescein Ingress to Corneal Epithelium

doi: 10.1167/iovs.63.2.31

Figure Lengend Snippet: Fluorescein punctate staining are the fluorescein-incorporated corneal epithelial cells of the superficial layer featured with decreased MUC1 expression. (A) The clinical or slit lamp view under cobalt blue light of the eyes of sham-operated rabbit model (lower panel) or dry eye rabbit model ( upper panel ) following fluorescein staining. (B) The fluorescein-stained corneal epithelia prepared as the frozen tissue sections were stained with MUC1 for immunohistochemistry ( brown , left panel ) and immunofluorescence ( middle and right panel ) analyses. For immunofluorescence, dual channels ( red and blue , middle panel ) or triple channels ( red , blue , and green , right panel ) were used. The cell nuclei were counterstained with hematoxylin ( purple , immunohistochemistry) or DAPI ( blue , immunofluorescence). (C) The frozen corneal epithelia section prepared from sham-operated and lacrimal gland-removed rabbits were immunostained with ZO-1 ( red ), nuclear counterstained with DAPI ( blue ), and the fluorescein ingress ( green ) were analyzed through a confocal microscopy. ( red and blue dual channels, middle panel ; red , blue , and green triple channels, right panel ).

Article Snippet: The expression of the truncated MUC1 was performed using MUC1 cDNA ORF Clone, human (HG12123-UT, SinoBiology) and Lipofectamine 3000 Reagent (Thermo Fisher Scientific) according to the manufacturer's instructions.

Techniques: Staining, Expressing, Immunohistochemistry, Immunofluorescence, Confocal Microscopy

Journal: Investigative Ophthalmology & Visual Science

Article Title: Transmembrane Mucin 1 Blocks Fluorescein Ingress to Corneal Epithelium

doi: 10.1167/iovs.63.2.31

Figure Lengend Snippet: The fluorescein intensity is reversely correlated with the level of MUC1 in HCECs cultured in MPM. (A) The HCECs that have been grown in KSFM were cultured in MPM for two or five days, and the levels of MUC1, MUC4, and MUC16 mRNA were examined. (B) Representative flow cytometric analyses of MUC1 expressions on cell membranes of HCECs cultured in KSFM ( left ) or MPM ( right ) were presented in contour plots. (C) Representative flow cytometric analyses of fluorescence intensity and MUC1 expression in HCECs cultured in KSFM ( left ) or MPM ( right ) followed by fluorescein staining were presented in contour plots. (D) The HCECs cultured in KSFM or MPM were stained with fluorescein, and the levels of fluorescence ( green ) and MUC1 ( red ) were examined using an immunofluorescence microscopy. Nuclei were stained with DAPI ( blue ). The images were obtained via a confocal microscopy.

Article Snippet: The expression of the truncated MUC1 was performed using MUC1 cDNA ORF Clone, human (HG12123-UT, SinoBiology) and Lipofectamine 3000 Reagent (Thermo Fisher Scientific) according to the manufacturer's instructions.

Techniques: Cell Culture, Fluorescence, Expressing, Staining, Immunofluorescence, Microscopy, Confocal Microscopy

Journal: Investigative Ophthalmology & Visual Science

Article Title: Transmembrane Mucin 1 Blocks Fluorescein Ingress to Corneal Epithelium

doi: 10.1167/iovs.63.2.31

Figure Lengend Snippet: Diquafosol treatments preferentially promotes the expression of MUC1 on cell membranes. (A) The HCECs were grown in a culture media containing 0, 0.5, or 1.0 mM diquafosol for four days, and the levels of MUC1, MUC4, and MUC16 mRNA of treated cells were examined using qRT-PCR. (B) Representative flow cytometric analyses of MUC1, MUC4, and MUC16 expression on cell membranes of HCECs treated with 1 mM diquafosol ( red ) or control ( gray ) were presented in overlaid histograms of signal intensity. (C) Representative flow cytometric analyses of fluorescence intensity and MUC1 expression in HCECs treated with sham ( left ) or 1 mM diquafosol ( right ) followed by fluorescein staining were presented in contour plots.

Article Snippet: The expression of the truncated MUC1 was performed using MUC1 cDNA ORF Clone, human (HG12123-UT, SinoBiology) and Lipofectamine 3000 Reagent (Thermo Fisher Scientific) according to the manufacturer's instructions.

Techniques: Expressing, Quantitative RT-PCR, Fluorescence, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Transmembrane Mucin 1 Blocks Fluorescein Ingress to Corneal Epithelium

doi: 10.1167/iovs.63.2.31

Figure Lengend Snippet: Knockdown of MUC1 leads to an increase in fluorescein ingress. (A) The mRNA levels of MUC1, MUC4, and MUC16 in HCECs separately transduced with two lentiviral-based shRNA vectors (shMUC1-1 or shMUC1-2) were examined using RT-PCR. The β-actin gene was used as an internal control. (B) Representative flow cytometric analyses of fluorescence intensity and MUC1 expressions in transduced HCECs followed by fluorescein staining were shown in contour plots.

Article Snippet: The expression of the truncated MUC1 was performed using MUC1 cDNA ORF Clone, human (HG12123-UT, SinoBiology) and Lipofectamine 3000 Reagent (Thermo Fisher Scientific) according to the manufacturer's instructions.

Techniques: Transduction, shRNA, Reverse Transcription Polymerase Chain Reaction, Fluorescence, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Transmembrane Mucin 1 Blocks Fluorescein Ingress to Corneal Epithelium

doi: 10.1167/iovs.63.2.31

Figure Lengend Snippet: The overexpression of MUC1 lacking the extracellular domain is unable to block fluorescein ingress. (A) Flow cytometric analyses of keratin 3, keratin 12, and vimentin expressions in HCECs cultured in KSFM ( gray ) or MPM ( red ) were presented in overlaid histograms of signal intensity. (B) Schematic diagrams of the wild-type and N-terminally truncated MUC1. (C) Representative flow cytometric analyses of fluorescence intensity and MUC1-C levels in HCECs transfected with the control vector ( left ) or the vector encoded with N-terminally truncated MUC1s ( right ) followed by fluorescein staining were shown in contour plots.

Article Snippet: The expression of the truncated MUC1 was performed using MUC1 cDNA ORF Clone, human (HG12123-UT, SinoBiology) and Lipofectamine 3000 Reagent (Thermo Fisher Scientific) according to the manufacturer's instructions.

Techniques: Over Expression, Blocking Assay, Cell Culture, Fluorescence, Transfection, Plasmid Preparation, Staining

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Knockout of Mucin 1 inhibits the proliferation, migration, and invasion of human MDA-MB-231 cells by blocking autophagy flow

doi: 10.1590/1414-431X2026e15075

Figure Lengend Snippet: Mucin 1 (MUC1) expression in human breast cancer. A , MUC1 expression levels in breast cancer (UALCAN website). B , MUC1 expression levels in different subtypes of breast cancer (UALCAN website). C , MUC1 protein expression in breast cancer (Human Protein Atlas database). D , MUC1 expression in human normal mammary epithelial cells MCF10A, TNBC cells (MDA-MB-231), and TNBC tissues (scale bar=100 μm). Data are reported as means±SE. **P<0.01, ***P<0.001, ****P<0.0001; two-tailed unpaired Student's t -test. BRCA: breast cancer gene; TCGA: The Cancer Genome Atlas; TNBC: triple-negative breast cancer.

Article Snippet: IHC data in the Human Protein Atlas database showed increased MUC1 protein expression in breast cancer tissue, but rarely expressed in normal breast tissue.

Techniques: Expressing, Two Tailed Test

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Knockout of Mucin 1 inhibits the proliferation, migration, and invasion of human MDA-MB-231 cells by blocking autophagy flow

doi: 10.1590/1414-431X2026e15075

Figure Lengend Snippet: Construction and validation the Mucin 1 (MUC1) knockout cell line 231-MUC1-KO. A , Sequencing results after MUC1 knockout were confirmed by the Snapgene software to identify the monoclonal cell line 231-MUC1-KO. B , RT-qPCR for MUC1 in MDA-MB-231 and 231-MUC1-KO cells. C , Western blot for MUC1 in MDA-MB-231 and 231-MUC1-KO cells. Data are reported as means±SE. ****P<0.0001; two-tailed unpaired Student's t -test.

Article Snippet: IHC data in the Human Protein Atlas database showed increased MUC1 protein expression in breast cancer tissue, but rarely expressed in normal breast tissue.

Techniques: Biomarker Discovery, Knock-Out, Sequencing, Software, Quantitative RT-PCR, Western Blot, Two Tailed Test

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Knockout of Mucin 1 inhibits the proliferation, migration, and invasion of human MDA-MB-231 cells by blocking autophagy flow

doi: 10.1590/1414-431X2026e15075

Figure Lengend Snippet: Effect of Mucin 1 (MUC1) knockout on the proliferation, migration, and invasion of MDA-MB-231 cells. A , MDA-MB-231 cell proliferation by EDU assay (scale bar=100 μm). B , MDA-MB-231 cell proliferation by colony formation assay. C , Migration and invasion of MDA-MB-231 cells by transwell assay (scale bar=150 μm). Data are reported as means±SE. **P<0.01, ***P<0.001, ****P<0.0001; two-tailed unpaired Student's t -test.

Article Snippet: IHC data in the Human Protein Atlas database showed increased MUC1 protein expression in breast cancer tissue, but rarely expressed in normal breast tissue.

Techniques: Knock-Out, Migration, EdU Assay, Colony Assay, Transwell Assay, Two Tailed Test

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Knockout of Mucin 1 inhibits the proliferation, migration, and invasion of human MDA-MB-231 cells by blocking autophagy flow

doi: 10.1590/1414-431X2026e15075

Figure Lengend Snippet: Effect of Mucin 1 (MUC1) knockout on autophagy flow in MDA-MB-231 cells. A , Expression of LC3 I, LC3 II, and P62 proteins by western blot. B , Ad-mCherry-GFP-LC3B-labeled LC3B protein assessed the patency of intracellular autophagy flow (scale bar=100 μm). Data are reported as means±SE. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; two-tailed unpaired Student's t -test. BafA1: Bafilomycin A1; Rapa: Rapamycin.

Article Snippet: IHC data in the Human Protein Atlas database showed increased MUC1 protein expression in breast cancer tissue, but rarely expressed in normal breast tissue.

Techniques: Knock-Out, Expressing, Western Blot, Labeling, Two Tailed Test

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Knockout of Mucin 1 inhibits the proliferation, migration, and invasion of human MDA-MB-231 cells by blocking autophagy flow

doi: 10.1590/1414-431X2026e15075

Figure Lengend Snippet: Effect of Mucin 1 (MUC1) knockout on lysosomes in MDA-MB-231 cells. A , Expression of SNAP29 and STXl7 by western blot. B , Expression of LAMP1 by western blot. C , pH was observed with red fluorescent probe Lysotracker Red (scale bar=100 μm). Data are reported as means±SE. **P<0.01, NS: not significant; two-tailed unpaired Student's t -test.

Article Snippet: IHC data in the Human Protein Atlas database showed increased MUC1 protein expression in breast cancer tissue, but rarely expressed in normal breast tissue.

Techniques: Knock-Out, Expressing, Western Blot, Two Tailed Test

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Knockout of Mucin 1 inhibits the proliferation, migration, and invasion of human MDA-MB-231 cells by blocking autophagy flow

doi: 10.1590/1414-431X2026e15075

Figure Lengend Snippet: Schematic diagram of the Mucin 1 (MUC1) oncogenic mechanism in breast cancer cell line MDA-MB-231. Knockout (KO) of MUC1 inhibits the proliferation, migration, and invasion of MDA-MB-231 cells by regulating the autophagic flow, and the possible mechanism may be related to the blocking of the autophagic flow caused by lysosomal damage.

Article Snippet: IHC data in the Human Protein Atlas database showed increased MUC1 protein expression in breast cancer tissue, but rarely expressed in normal breast tissue.

Techniques: Knock-Out, Migration, Blocking Assay

Journal: Plant biotechnology journal

Article Title: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice.

doi: 10.1111/j.1467-7652.2011.00614.x

Figure Lengend Snippet: Figure 1 magnICONTM viral vector transient expression system. (a) Dia- grammatic representation of magnICONTM pro-vector modules used in this study. Hsp81.1 represents the Arabidopsis heat shock protein 81.1 promoter; Integrase, Streptomyces phage PhiC31 integrase; AttP and AttB, PhiC31 integrase recombination sites; Act2, Arabidopsis actin 2 promoter; RdRP, RNA-dependent RNA polymerase; MP, movement pro- tein; LTB, synthetic LTB gene; LTB-G-MUC1, LTB gene with the GPGP linker and the MUC1 tandem repeat; LTB-G-MUC1-6His, LTB-G-MUC1 gene with a histidine tag on the C-terminus of MUC1; GFP, green fluo- rescent protein; LTBHis, LTB gene with the SGPS linker and a histidine tag, 3¢NTR, 3¢ untranslated region; NosT, nopaline synthase terminator, Int, intron; LB and RB, left and right borders of the T-DNA region. (b) Northern analysis of Nicotiana benthamiana infiltrated with magnI- CONTM pro-vector modules, for the detection of LTB-MUC1 mRNA. Lanes: 1–2, N. benthamiana infiltrated with pICH-LM1; 3, N. benthami- ana infiltrated with pICH-LTB; W.T., noninfiltrated N. benthamiana control. Bottom panel is the methylene blue-stained membrane for loading control: 10 lg of total RNA was loaded for each sample.

Article Snippet: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice Julia Pinkhasov1, M. Lucrecia Alvarez1, M. Manuela Rigano1, Khanrat Piensook1, Dalia Larios1, Martin Pabst2, Josephine Grass2, Pinku Mukherjee3, Sandra J. Gendler3, Amanda M. Walmsley1 and Hugh S. Mason1,* 1The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA 2Department of Chemistry, Universitaet fuer Bodenkultur, Wien, Austria 3Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Mayo Clinic, Scottsdale, AZ, USA Received 16 November 2009; revised 27 January 2011; accepted 17 February 2011.

Techniques: Plasmid Preparation, Expressing, Northern Blot, Control, Staining, Membrane

Journal: Plant biotechnology journal

Article Title: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice.

doi: 10.1111/j.1467-7652.2011.00614.x

Figure Lengend Snippet: Figure 2 Time course of LTB-MUC1 expression in magnICONTM infil- trated Nicotiana benthamiana. (a) Expression of GFP and LTB-MUC1 in N. benthamiana 7, 9 and 11 days postinfiltration (dpi). Slight necrosis is apparent in LTB-MUC1-expressing leaves 9 dpi, with complete necrosis at infiltration site by day 11. Ganglioside capture ELISA was used to quantify LTB (b) and MUC1 (c) expression at 7, 9 and 11 days postinfil- tration. Expression levels reported in micrograms per gram fresh weight (lg ⁄ g f.w.). Data are means of three independent leaf infiltrations ±SD.

Article Snippet: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice Julia Pinkhasov1, M. Lucrecia Alvarez1, M. Manuela Rigano1, Khanrat Piensook1, Dalia Larios1, Martin Pabst2, Josephine Grass2, Pinku Mukherjee3, Sandra J. Gendler3, Amanda M. Walmsley1 and Hugh S. Mason1,* 1The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA 2Department of Chemistry, Universitaet fuer Bodenkultur, Wien, Austria 3Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Mayo Clinic, Scottsdale, AZ, USA Received 16 November 2009; revised 27 January 2011; accepted 17 February 2011.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Plant biotechnology journal

Article Title: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice.

doi: 10.1111/j.1467-7652.2011.00614.x

Figure Lengend Snippet: Figure 3 Western blot analysis of metal affinity-purified magnICONTM-expressed LTB-6His and LTB-MUC1-6His. (a) Anti-LTB antibody was used to detect LTB-6His (lane LTB) and LTB-MUC1-6His (lane LM1). The LTB lane showed the LTB monomer at the expected molecular mass of 12 kDa as well as a possible dimer at 24 kDa. The LM1 lane showed the expected size of the LTB-MUC1-6His monomer at 16 kDa, as well as two higher molecular mass proteins at approximately 18 and 20 kDa. The protein detected at approximately 32 kDa may represent the dimer for the LTB-MUC1-6His. (b) Nickel-conjugated HRP was used to detect LTB-6His (lane LTB) and LTB-MUC1-6His (lane LM1). The LTB lane showed the His-tagged LTB monomer at 12 kDa. The LM1 lanes only showed the higher molecular mass LTB-MUC1-6His proteins, approximately 18 and 20 kDa, but not the 16-kDa protein. (c) Anti-MUC1 BC2 monoclonal antibody was used to detect LTB-6His (lane LTB) and LTB-MUC1-6His (lane LM1). No MUC1 protein was detected in the LTB lane. Only the approximately 18 and 20 kDa of LTB-MUC1-6His were detected by the MUC1-specific antibody.

Article Snippet: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice Julia Pinkhasov1, M. Lucrecia Alvarez1, M. Manuela Rigano1, Khanrat Piensook1, Dalia Larios1, Martin Pabst2, Josephine Grass2, Pinku Mukherjee3, Sandra J. Gendler3, Amanda M. Walmsley1 and Hugh S. Mason1,* 1The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA 2Department of Chemistry, Universitaet fuer Bodenkultur, Wien, Austria 3Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Mayo Clinic, Scottsdale, AZ, USA Received 16 November 2009; revised 27 January 2011; accepted 17 February 2011.

Techniques: Western Blot

Journal: Plant biotechnology journal

Article Title: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice.

doi: 10.1111/j.1467-7652.2011.00614.x

Figure Lengend Snippet: Figure 4 Glycosylation analysis of LTB-Muc1-6His. (a) Coomassie-stained SDS–PAGE gel of purified LTB-6His (lanes 1 and 2) and LTB-MUC1-6His (lanes 4 and 5) protein fractions. Gel bands at approximately 12 kDa (LTB-6His) and 16, 18 and 20 ⁄ 22 kDa (LTB-Muc1-6His) were excised and hydrolysed with TFA to release monosaccharides. (b) RP-HPLC of LTB-MUC1-6His shows a distinct peak for arabinose from the 20 ⁄ 22 and 18-kDa molecular weight fractions, and a shorter peak from the 16-kDa fraction. An excised piece from the gel served as a negative control (Gel blank). (c) RP-HPLC of LTB-6His does not show any monosaccharide peaks, suggesting the LTB is not glycosylated. The standard contained the following glycans: GlcN, Gluco- samine; ManN, Mannosamine; GalN, Galactosamine; Gal, Galactose; Man, Mannose; Glc, Glucose; Ara, Arabinose; Xyl, Xylose. Gel blank was used as a negative control.

Article Snippet: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice Julia Pinkhasov1, M. Lucrecia Alvarez1, M. Manuela Rigano1, Khanrat Piensook1, Dalia Larios1, Martin Pabst2, Josephine Grass2, Pinku Mukherjee3, Sandra J. Gendler3, Amanda M. Walmsley1 and Hugh S. Mason1,* 1The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA 2Department of Chemistry, Universitaet fuer Bodenkultur, Wien, Austria 3Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Mayo Clinic, Scottsdale, AZ, USA Received 16 November 2009; revised 27 January 2011; accepted 17 February 2011.

Techniques: Glycoproteomics, Staining, SDS Page, Molecular Weight, Negative Control

Journal: Plant biotechnology journal

Article Title: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice.

doi: 10.1111/j.1467-7652.2011.00614.x

Figure Lengend Snippet: Figure 5 Pronase peptide analysis of LTB-MUC1-6His by MS and MS ⁄ MS. (a) Amino acid sequence of LTB-MUC1-6His. Numbers at left indicate the amino acid positions from the N-terminus. The amino acid sequence GSTAPPAHG that is glycosylated with arabinose is bold ⁄ italic ⁄ dotted underlined; the tri-arabinan is found on the double proline site within this sequence. Other prolines are bold ⁄ underlined, and double prolines are bold ⁄ double underlined. (b) MS and MS ⁄ MS analysis of pronase digestion products. LTB-MUC1-6His gel fractions were pooled and subjected to digestion with pronase. One small peptide with the amino acid sequence GSTAPPAHG comprising proline residues 113 and 114 was identified with three arabinose residues attached. This glycopeptide was only found in LTB-MUC1-6His samples but not in LTB-6His.

Article Snippet: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice Julia Pinkhasov1, M. Lucrecia Alvarez1, M. Manuela Rigano1, Khanrat Piensook1, Dalia Larios1, Martin Pabst2, Josephine Grass2, Pinku Mukherjee3, Sandra J. Gendler3, Amanda M. Walmsley1 and Hugh S. Mason1,* 1The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA 2Department of Chemistry, Universitaet fuer Bodenkultur, Wien, Austria 3Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Mayo Clinic, Scottsdale, AZ, USA Received 16 November 2009; revised 27 January 2011; accepted 17 February 2011.

Techniques: Sequencing, Glycoproteomics

Journal: Plant biotechnology journal

Article Title: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice.

doi: 10.1111/j.1467-7652.2011.00614.x

Figure Lengend Snippet: Figure 6 Analysis of Ara-Hyp species in LT-MUC1-6His. (a) Carbon- LC-ESI-MS analysis of LTB-MUC1-6His after alkaline degradation. A high percentage of the prolines as hydroxyprolines (33%) or glycosylated hydroxyprolines, mainly with a tri-arabinan (11% of total prolines) were observed. Hyp and glyco-Hyp give two peaks containing different con- formers. (b) Quantitative comparison of the content of Proline (Pro), hydroxyproline (Hyp) and glycosylated hydroxyproline between LTB-6His and LTB-MUC1-6His. Both samples contained proline and hydroxypro- line. LTB-MUC1-6His contained more hydroxyproline than LTB-6His and contained Ara-Hyp glycopeptides, mainly with three arabinose residues. Ara2, Ara3 and Ara4 designate a Hyp with 2, 3 and 4 Ara residues, respectively.

Article Snippet: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice Julia Pinkhasov1, M. Lucrecia Alvarez1, M. Manuela Rigano1, Khanrat Piensook1, Dalia Larios1, Martin Pabst2, Josephine Grass2, Pinku Mukherjee3, Sandra J. Gendler3, Amanda M. Walmsley1 and Hugh S. Mason1,* 1The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA 2Department of Chemistry, Universitaet fuer Bodenkultur, Wien, Austria 3Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Mayo Clinic, Scottsdale, AZ, USA Received 16 November 2009; revised 27 January 2011; accepted 17 February 2011.

Techniques: Comparison

Journal: Plant biotechnology journal

Article Title: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice.

doi: 10.1111/j.1467-7652.2011.00614.x

Figure Lengend Snippet: Figure 7 Parenteral immunization of MUC1.Tg mice with purified magnICONTM generated LTB- MUC1-6His. (a) Serum analysis for MUC1-specific IgG in MUC1.Tg mice treated with: phosphate buffer saline vehicle control (PBS); 100 lg CpG ODN (CpG); 100 lg CpG + LTB-MUC1-6His (LM1CpG). Serum samples were derived from blood drawn 1 week after each immunization (days 0, 7 and 14) and boost (day 35). (b) Serum MUC1-specific IgG1 and IgG2a (c) in MUC1.Tg mice treated with: PBS; 100 lg CpG ODN (CpG); LTB-MUC1-6His (LTB-MUC1); 100 lg CpG + LTB- MUC1-6His (CpG + LTB-MUC1). Anti-MUC1 titres (IgG, IgG1 and IgG2a) in mice treated with CpG + LTB-MUC1 are significantly higher (P < 0.05) than the PBS control group. Individual mouse data are shown (n = 3–5 mice per group).

Article Snippet: Recombinant plant-expressed tumour-associated MUC1 peptide is immunogenic and capable of breaking tolerance in MUC1.Tg mice Julia Pinkhasov1, M. Lucrecia Alvarez1, M. Manuela Rigano1, Khanrat Piensook1, Dalia Larios1, Martin Pabst2, Josephine Grass2, Pinku Mukherjee3, Sandra J. Gendler3, Amanda M. Walmsley1 and Hugh S. Mason1,* 1The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA 2Department of Chemistry, Universitaet fuer Bodenkultur, Wien, Austria 3Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Mayo Clinic, Scottsdale, AZ, USA Received 16 November 2009; revised 27 January 2011; accepted 17 February 2011.

Techniques: Generated, Saline, Control, Derivative Assay