Journal: bioRxiv

Article Title: The Ry sto immune receptor recognizes a broadly conserved feature of potyviral coat proteins

doi: 10.1101/2021.05.20.444601

Figure Lengend Snippet: (a) Phylogeny of Potyviridae CPs based on the amino acid sequences aligned using ClustalW and calculated with Mega7. Cloned sequences are marked in red. (b) Sequence logo analysis of CPs of 65 potyviruses depicting the conservation of the predicted core region, including the S 126 _R 157 _D 201 ssRNA pocket binding site. (c) CPs of various potyviruses trigger Ry sto -mediated immunity. Ry sto or control tobacco plants were agroinfiltrated with multiple potyviral CPs: PRSV, papaya ringspot virus; PVM, potato virus M; SMV, soybean mosaic virus; PepSMV, pepper severe mosaic virus; CBMNV, bean common mosaic necrosis virus; MDMV, maize dwarf mosaic virus; TuMV, turnip mosaic virus; PSbMV, pea-seed born mosaic virus; SPFMV, sweet potato feathery mottle virus; or PPV, plum pox virus. All of them elicit Ry sto -dependent immune response. The photographs were obtained at 72 hours postinfiltration (hpi). Empty vector was used as a negative control (CTRL). (d) In planta accumulation of the potyviral CPs. For anti-FLAG immunoblots of viral CPs total protein extracts were prepared from leaves tested at 3 days after agroinfiltration. Staining of RuBisCo with Ponceau S was used as a loading control.

Article Snippet: We site-inoculated leaves of transgenic Ry sto and control tobacco plants with a severe PPV isolate (DSMZ No.: PV-0001).

Techniques: Clone Assay, Sequencing, Binding Assay, Control, Virus, Plasmid Preparation, Negative Control, Western Blot, Staining

Journal: bioRxiv

Article Title: The Ry sto immune receptor recognizes a broadly conserved feature of potyviral coat proteins

doi: 10.1101/2021.05.20.444601

Figure Lengend Snippet: (a) PPV-induced cell death development visualised by trypan blue staining in Ry sto -expressing or control, tobacco plants locally inoculated with a severe PPV isolate (PV-001, DSMZ). At 7 dpi, Ry sto plants exhibited a local cell death, whereas controls remained symptomless. (b) Ry sto confers resistance to short-distance PPV movement. Ry sto transgenic and control plants were site-infected with different PPV isolates (PV-0001 or PV-2233, DSMZ). At 7 and 14 dpi, samples from consecutive leaf zones were harvested and PPV proteins were detected using anti-PPV antibodies. Staining of RuBisCo with Ponceau S was used as a loading control. (c) Leaves of or Ry sto -expressing and control plants were agroinfiltrated with PPV tagged with GFP. The viral spread was monitored by GFP-fluorescence. Photographs were obtained at 14 dpi under blue light (488 nm).

Article Snippet: We site-inoculated leaves of transgenic Ry sto and control tobacco plants with a severe PPV isolate (DSMZ No.: PV-0001).

Techniques: Staining, Expressing, Control, Transgenic Assay, Infection, Fluorescence

Journal: Nature Communications

Article Title: Mutant ASXL1 induces age-related expansion of phenotypic hematopoietic stem cells through activation of Akt/mTOR pathway

doi: 10.1038/s41467-021-22053-y

Figure Lengend Snippet: a The experimental design for treatment with an mTOR inhibitor rapamycin in aged mice. Aged Vav-Cre ASXL1-MT KI mice and age-matched control mice were treated with rapamycin intraperitoneally (4 mg/kg/day) every other day for 8 weeks. At the end of administrations, end-point analyses of bone marrow cells were conducted. b , c The frequency of LT-HSCs in bone marrow MNCs ( n = 6 (Control-Vehicle), 7 (Control-Rapamycin), 5 (ASXL1-MT-Vehicle), and 7 (ASXL1-MT-Rapamycin)). Representative FACS plot ( b ) and summarized data ( c ) are shown. d Cell cycle analysis with Ki-67/DAPI staining of LT-HSCs ( n = 7 (Young control), 6 (Control-Vehicle), 7 (Control-Rapamycin), 5 (ASXL1-MT-Vehicle), and 7 (ASXL1-MT-Rapamycin)). e Apoptosis analyses of LT-HSCs ( n = 5 (Control-Vehicle), 7 (Control-Rapamycin), 5 (ASXL1-MT-Vehicle), and 7 (ASXL1-MT-Rapamycin)). f The experimental design for treatment with an Akt inhibitor perifosine in aged mice. Aged Vav-Cre ASXL1-MT KI mice and age-matched control mice were treated with a single oral dose (36 mg/kg) of perifosine ( n = 5). g 16 h after administration, cell cycle analyses with Ki-67/DAPI staining of LT-HSCs were performed. Statistical significances are assessed by one-way ANOVA with Tukey–Kramer’s post-hoc test. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

Article Snippet: Perifosine (Chemscene LLC, CS-0209) was dissolved in sterile water, and 36 mg/kg was administered orally 16 h before analyses.

Techniques: Control, Cell Cycle Assay, Staining