Journal: Frontiers in Immunology

Article Title: Characterization of intestinal mononuclear phagocyte subsets in young ruminants at homeostasis and during Cryptosporidium parvum infection

doi: 10.3389/fimmu.2024.1379798

Figure Lengend Snippet: List of antibodies.

Article Snippet: Anti-ovine MHC II-RPE , MHCII , Mouse IgG1 , 28.1 , Bio-Rad , Cattle , Datasheet MCA2225PE.

Techniques:

Journal: The Plant Cell

Article Title: Strigolactone Promotes Degradation of DWARF14, an α/β Hydrolase Essential for Strigolactone Signaling in Arabidopsis ^[W]

doi: 10.1105/tpc.114.122903

Figure Lengend Snippet: Phenotypes of d14-seto Single and Double mutants. (A) Arabidopsis branching structure. (B) Close-up of mature wild-type Col-0 (left) and seto5 (here termed d14-seto) (right) rosettes showing their lateral shoot phenotype. Bar = 1 cm. (C) Number of primary rosette branches (RI) of wild-type and d14-seto plants. (D) Number of secondary branches (RII+CII) relative to the number of primary branches (RI+CI). (E) Height of the main inflorescence of the same set of plants. (F) Developmental stages of buds in the axils of cotyledons (C1 and C2) and rosette leaves (L1 to L10) of wild-type (top) and d14-seto (bottom) individuals. R, reproductive stage, V1 to V3, vegetative stages; LP, leaf primordium stages; M, meristem; E, empty axil. Developmental stages are as defined (Aguilar-Martínez et al., 2007) (n = 10). (G) Flowering time, expressed as number of leaves, of lateral inflorescences of wild-type, d14-seto, and brc1-2 mutants and d14-seto brc1-2 double mutants. −1, uppermost RI; +1 and +2, first and second basal-most CI branches. (H) and (I) Number of RI branches (H) and height of the main inflorescence (I) of d14-seto max2-1 double mutants. Asterisks denote significant differences in Student’s t tests (P < 0.0001). Letters denote significant differences in one-way ANOVA test (Tukey test P < 0.05). Data shown as mean ± se (n = 20).

Article Snippet: For fine mapping, 400 additional F2 plants were used to assess linkage iteratively between seto5 and molecular markers designed according to the polymorphisms between Landsberg erecta and Col-0 described at the Monsanto Arabidopsis Polymorphism Collection database ( http://www.arabidopsis.org ).

Techniques:

Journal: The Plant Cell

Article Title: Strigolactone Promotes Degradation of DWARF14, an α/β Hydrolase Essential for Strigolactone Signaling in Arabidopsis ^[W]

doi: 10.1105/tpc.114.122903

Figure Lengend Snippet: The d14-seto Protein. (A) Sequence alignment of the Arabidopsis D14 segment comprising the Pro169Leu mutation, with ortholog sequences petunia DAD2 (Hamiaux et al., 2012), rice D14 (Arite et al., 2009; Gao et al., 2009; Liu et al., 2009; Hamiaux et al., 2012), paralog KAI2 (Waters et al., 2012b), and related bacterial protein RsbQ (Brody et al., 2001). Red arrow indicates Pro-169 and corresponding amino acid Ser-168 in KAI2. Asterisks indicate residues Met-166 and Ile-169, which undergo conformational changes in KAI2 after KAR1 binding (Guo et al., 2013b). Horizontal red bars indicate the position of two of the KAI2 cap α-helices (Kagiyama et al., 2013). (B) Logos of SDPs that differ in D14 (top) and KAI2 (bottom) ortholog sequences. Numbering corresponds to D14 and KAI2 protein sequences. Pro-169 and Ser-168 are shown in red. Hydrophobic residues are indicated in black, polar residues in green, and Gly and Pro in yellow. Letter size represents percentage of conservation within protein classes. (C) Front view of D14 (PDB:4ih4) and KAI2 (PDB:3w06) structural alignment. Helical caps of D14 and KAI2 are highlighted in salmon pink and yellow, respectively. Active site residues are in blue, D14 Pro-169 is in red, and KAI2 S168 is in green. (D) Close-up view and side chain superposition of wild-type D14 P169 (red), mutant Leu-169 (blue), and KAI2 Ser-168 (green). Note that the Pro-169 side chain is exposed to the solvent and that KAI2 loop (yellow) is longer than that of D14. (E) D14 structure in surface representation. Residues corresponding to residues in KAI2 that undergo side-chain movement after KAR1 binding are labeled and highlighted in red, Pro-169 in purple and cap domain in pink.

Article Snippet: For fine mapping, 400 additional F2 plants were used to assess linkage iteratively between seto5 and molecular markers designed according to the polymorphisms between Landsberg erecta and Col-0 described at the Monsanto Arabidopsis Polymorphism Collection database ( http://www.arabidopsis.org ).

Techniques: Sequencing, Mutagenesis, Binding Assay, Solvent, Labeling

Journal: The Plant Cell

Article Title: Strigolactone Promotes Degradation of DWARF14, an α/β Hydrolase Essential for Strigolactone Signaling in Arabidopsis ^[W]

doi: 10.1105/tpc.114.122903

Figure Lengend Snippet: D14 Promoter Activity and D14 Protein Distribution during Arabidopsis Development. GUS histochemical activity of Arabidopsis D14pro:GUS ([A] to [Q]) and D14pro:D14:GUS ([R] to [W]) transgenic plants. (A) Five-day-old transgenic seedlings. The plant on the right, more advanced in development, shows expression in the root more restricted to the vascular cylinder (arrowheads) than that of the less developmentally advanced (left). (B) Ten-day-old seedling with GUS activity in the vascular tissue of the hypocotyl (arrowhead). (C) Eighteen-day-old vegetative rosette. (D) Young rosette leaf from plant in (C). (E) Mature cauline leaf from 30-day-old plant. (F) Stem of the main inflorescence showing a gradient of GUS activity with a maximum near the apex. (G) Bud in the axil of a young rosette leaf. (H) Bud in the axil of a mature rosette leaf. (I) Detail of a rosette leaf surface. Note the separation between the xylem (white) bundle (black arrow) and the phloem (blue) bundle expressing GUS (blue arrow). (J) Main inflorescence. GUS accumulates in the apical-most stem region and in flower pedicels. (K) Close-up of a developing flower. Signal in the style is indicated (white arrow). (L) Root tip. (M) to (O) The 3-μm transverse plastic-embedded sections of root similar to that in (L). (M) Distal section showing GUS staining in procambium cells. (N) GUS is excluded from xylem cells (X). (O) More proximal section showing promoter activity in phloem cells (arrows). (P) Transverse plastic-embedded section of a stem internode of the primary inflorescence. (Q) Close-up of a section similar to that shown in (M), with cortex cells but not epidermis cells expressing GUS. Notice the stronger signal in the vascular bundle sector flanked by the arrows in (P) and (Q). (R) Root tip similar to that in (L). D14:GUS is present in the root tip. (S) to (U) The 3-μm transverse plastic-embedded sections of root tips. (S) shows the meristematic zone, and (T) and (U) are sections similar to those in (N) and (O). GUS signal is widespread in (S) and (T) and accumulates in the epidermis, cortex, and phloem (arrowheads) in (U). (V) and (W) Stem transverse plastic-embedded sections comparable to those in (P) to (Q). GUS is detectable throughout the cortex, epidermis, and phloem (arrowheads). (X) GFP fluorescence image (top) and fluorescence merged with bright-field image (bottom) of a transgenic D14pro:D14:GFP root. (Y) to (Z’) Leaf (Y), hypocotyl (Z), and root (Z’) cells of CaMV35Spro:D14:GFP transgenic plants. GFP is detected in nucleus and cytoplasm. Bars = 1 mm in (A), (D) to (F), and (K), 500 μm in (L), 200 μm in (B), (M), and (P), 100 μm in (G) and (N), 50 μm in (H) to (J), (Q), and (Z), 15 μm in (Y) and (Z’).

Article Snippet: For fine mapping, 400 additional F2 plants were used to assess linkage iteratively between seto5 and molecular markers designed according to the polymorphisms between Landsberg erecta and Col-0 described at the Monsanto Arabidopsis Polymorphism Collection database ( http://www.arabidopsis.org ).

Techniques: Activity Assay, Transgenic Assay, Expressing, Staining, Fluorescence

Journal:

Article Title: Characterization of Tocopherol Cyclases from Higher Plants and Cyanobacteria. Evolutionary Implications for Tocopherol Synthesis and Function 1

doi: 10.1104/pp.103.024257

Figure Lengend Snippet: HPLC analysis of tocopherols in wild-type and mutant Arabidopsis, maize, and Synechocystis sp. PCC6803. Tocopherols present in Arabidopsis, maize, and Synechocystis sp. PCC6803 lipid extracts were separated by normal phase HPLC and detected using a fluorescence detector with 290-nm excitation and 325-nm emission. Tocol, a synthetic tocopherol, was used as an internal recovery standard. A, Arabidopsis leaf tissue: solid line, Columbia wild type; dotted line, vte1-1; gray line, vte1-2. B, Maize leaf tissue: solid line, wild type; dotted line, sxd1. C, Synechocystis sp. PCC6803: solid line, wild type; dotted line, Δslr1737 insertional mutant; gray line, SXD1 expressed in the Δslr1737 insertional mutant. Retention times of α-, β-, δ-, and γ-tocopherol and tocol were determined by HPLC analysis of tocopherol standards. LU, Luminescence units.

Article Snippet: PCR-based markers were designed using INDEL or SNP from the Cereon Arabidopsis Polymorphism and Landsberg erecta Sequence Collection (Cereon Genomics LLC, Cambridge, MA; Jander et al., 2002 ).

Techniques: Mutagenesis, Fluorescence

Journal:

Article Title: Characterization of Tocopherol Cyclases from Higher Plants and Cyanobacteria. Evolutionary Implications for Tocopherol Synthesis and Function 1

doi: 10.1104/pp.103.024257

Figure Lengend Snippet: HPLC analysis of the prenyl quinones from wild-type and mutant Arabidopsis, maize, and Synechocystis sp. PCC6803. Lipids were extracted from Arabidopsis, maize, and Synechocystis sp. PCC6803, and total prenyl quinines were isolated by thin-layer chromatography (TLC) and then analyzed by normal phase HPLC (see “Materials and Methods”) A, Arabidopsis. Solid line, Columbia wild type; dotted line, vte1-1; gray line, vte1-2. B, Maize. Solid line, Wild type; dotted line, sxd1. C, Synechocystis sp. PCC6803. Solid line, Wild type; dotted line, Δslr1737 insertional mutant; gray line, SXD1cDNA expressed in the Δslr1737 mutant background. Insets, Spectra of the peak labeled DMPBQ. Phyllo, Phylloquinone; PQ, Plastoquinone.

Article Snippet: PCR-based markers were designed using INDEL or SNP from the Cereon Arabidopsis Polymorphism and Landsberg erecta Sequence Collection (Cereon Genomics LLC, Cambridge, MA; Jander et al., 2002 ).

Techniques: Mutagenesis, Isolation, Thin Layer Chromatography, Labeling

Journal:

Article Title: Characterization of Tocopherol Cyclases from Higher Plants and Cyanobacteria. Evolutionary Implications for Tocopherol Synthesis and Function 1

doi: 10.1104/pp.103.024257

Figure Lengend Snippet: HPLC analysis of seed tocopherols in wild-type Arabidopsis, vte1-1, and vte1-2. Total seed lipids were extracted, and the tocopherols present were separated by reverse phase HPLC and detected using a fluorescence detector; 290-nm excitation and 325-nm emission. Tocol, a synthetic tocopherol, was used as an internal recovery standard. Solid line, Columbia wild type; dotted line, vte1-1; gray line, vte1-2. Retention times of α-, δ-, and γ-tocopherol and tocol were determined by HPLC analysis of tocopherol standards.

Article Snippet: PCR-based markers were designed using INDEL or SNP from the Cereon Arabidopsis Polymorphism and Landsberg erecta Sequence Collection (Cereon Genomics LLC, Cambridge, MA; Jander et al., 2002 ).

Techniques: Fluorescence

Journal:

Article Title: Characterization of Tocopherol Cyclases from Higher Plants and Cyanobacteria. Evolutionary Implications for Tocopherol Synthesis and Function 1

doi: 10.1104/pp.103.024257

Figure Lengend Snippet: TC activity of proteins expressed in E. coli. E. coli cell lysates from cells overexpressing the empty pET vector or pET engineered to express TC proteins from Arabidopsis, maize, and Synechocystis sp. PCC6803 were incubated with radiolabeled 2,3-methyl-6-phytyl-1,4-benzonequinol (3 methyl 14C) for 4 h as described in “Materials and Methods.” Total lipids were extracted, separated by TLC, and radiolabeled products were detected by phosphor imager analysis. Products were identified by comigration with standards. The 14C incorporation into γ-tocopherol was quantified densitometrically and expressed as pixels per microgram of total protein.

Article Snippet: PCR-based markers were designed using INDEL or SNP from the Cereon Arabidopsis Polymorphism and Landsberg erecta Sequence Collection (Cereon Genomics LLC, Cambridge, MA; Jander et al., 2002 ).

Techniques: Activity Assay, Plasmid Preparation, Incubation

Journal:

Article Title: Characterization of Tocopherol Cyclases from Higher Plants and Cyanobacteria. Evolutionary Implications for Tocopherol Synthesis and Function 1

doi: 10.1104/pp.103.024257

Figure Lengend Snippet: Analysis of Glc, Suc, and starch in wild-type Arabidopsis and vte1-1 Glc, Suc, and starch levels were analyzed spectrophotometrically using the enzyme-coupled assays described in “Materials and Methods.” Glc and Suc are expressed as nanomoles per milligram fresh wt ( n = 4).

Article Snippet: PCR-based markers were designed using INDEL or SNP from the Cereon Arabidopsis Polymorphism and Landsberg erecta Sequence Collection (Cereon Genomics LLC, Cambridge, MA; Jander et al., 2002 ).

Techniques:

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Intestinal CD8 alpha alpha and CD8 alpha beta intraepithelial lymphocytes are thymus derived and exhibit subtle differences in TCR beta repertoires.

doi: 10.4049/jimmunol.165.12.6716

Figure Lengend Snippet: FIGURE 1. CD8aa cells are absent from the thymus. Cytofluorometry of E14 and adult thymocytes and adult gut using anti-CD8a and CD8b Abs 11-39 and EP-42, respectively, and anti-mouse IgG-specific Abs coupled to FITC or PE. Arrows point to the locations of CD8aa cells

Article Snippet: For three-color analysis we used mouse-anti-TCR Vb1 mAb coupled to biotin (TCR2, Southern Biotechnology, Birmingham, AL), mouse antiCD8b mAb (Ep42, an IgG2a, gift from M. Ratcliffe, Montreal, Canada), and mouse anti-CD8a mAbs (11-39, an IgG1 recognizing all polymorhic forms; 11-7 and 11-13, both IgG1 recognizing only CD8 of the strain H.B15.H12; (32)).

Techniques:

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Intestinal CD8 alpha alpha and CD8 alpha beta intraepithelial lymphocytes are thymus derived and exhibit subtle differences in TCR beta repertoires.

doi: 10.4049/jimmunol.165.12.6716

Figure Lengend Snippet: FIGURE 2. Identification of two chicken strains congenic for the CD8 a-chain. H.B15.H7 and H.B15.H12 strains express different CD8a alleles. A, Cytofluorometry of adult H7 and H12 chicken thymocytes using anti-CD8a Abs 11-13 and 11-39. Note that Ab 11-13 does not recognize CD8a on thy- mocytes of H7 animals. B, mAbs 11-13 and 11-39 immunoprecipitate the CD8 a-chain. The thymocytes of a 3-wk-old H12 chicken were 125I labeled. The lysate was precipitated with mAbs 11-13 and 11-39. Immunoprecipitates were analyzed by SDS-PAGE on a 10% gel under reducing conditions. The mo- lecular mass standards are indicated on the left. C, mAb 11-13 recognizes the allotypic CD8 a-chain. COS-7 cells were transfected with pCDM8 plasmids carrying CD8a from the inbred chicken lines H.B15.H7 (a) or H.B15.H12 (b), respectively. The cells were then stained with the CD8a allotypic mAb 11-13. Only H12 CD8 was recognized. Staining of the cells with mAb 11-39 served as a positive control. Abs were detected with HRP-conjugated rabbit anti- mouse-Ig.

Article Snippet: For three-color analysis we used mouse-anti-TCR Vb1 mAb coupled to biotin (TCR2, Southern Biotechnology, Birmingham, AL), mouse antiCD8b mAb (Ep42, an IgG2a, gift from M. Ratcliffe, Montreal, Canada), and mouse anti-CD8a mAbs (11-39, an IgG1 recognizing all polymorhic forms; 11-7 and 11-13, both IgG1 recognizing only CD8 of the strain H.B15.H12; (32)).

Techniques: Labeling, SDS Page, Transfection, Staining, Positive Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Intestinal CD8 alpha alpha and CD8 alpha beta intraepithelial lymphocytes are thymus derived and exhibit subtle differences in TCR beta repertoires.

doi: 10.4049/jimmunol.165.12.6716

Figure Lengend Snippet: FIGURE 3. Embryonic TCRgd1 CD8aa2 thymocytes differentiate into TCRgd1 CD8aa1 iIELs. E14 H12 thymocytes (2 3 107) were in- jected i.v. into E16 H7 recipient embryos. The iIELs were analyzed 18 days after injection by cytofluorometry. CD8aa1 donor cells were detected by Ab 11-13, which specifically recognizes the H12 CD8 a-chain (donor). Total CD8a1 iIELs, including both host and donor cells from the same animal, were detected with Ab 11-39 recognizing donor and host CD8a1

Article Snippet: For three-color analysis we used mouse-anti-TCR Vb1 mAb coupled to biotin (TCR2, Southern Biotechnology, Birmingham, AL), mouse antiCD8b mAb (Ep42, an IgG2a, gift from M. Ratcliffe, Montreal, Canada), and mouse anti-CD8a mAbs (11-39, an IgG1 recognizing all polymorhic forms; 11-7 and 11-13, both IgG1 recognizing only CD8 of the strain H.B15.H12; (32)).

Techniques: Injection