Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) Body size and appearance in Ten1 hom mice at P23. ( B ) Survival analysis (Kaplan-Meier: P < 0.0001; n WT/hom: 25/29). ( C ) Body weight at P0.5 and P23 (unpaired t test: P < 0.0001 each; n P0.5 WT/hom: 17/18, P23 WT/hom: 21/15). ( D ) μCT imaging for skeleton analysis.

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Imaging

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) Telomere length by qPCR in the cerebrum, cerebellum, liver, and lung of Ten1 hom animals at 23 days of age. ( B ) Telomere length by qPCR at P5 and P23 in the skin of Ten1 WT and hom animals. ( C ) Representative images of Q-FISH immunofluorescence staining in the cerebellum, liver, lung, and small intestine at P23. The insets show magnification images of WT (left) and hom (right) cells below each tissue. Violin plots showing the ( D ) number of telomeric foci per nucleus and ( E ) mean nuclear telomeric intensity determined by Q-FISH in the cerebellum, liver, lung, and small intestine of Ten1 WT and hom P23 animals ( n WT/hom: 5/5). a.u.f., arbitrary units of fluorescence. Three to four images corresponding to different areas of the tissues were analyzed from each mouse. The median and the upper and lower quartiles of each distribution are indicated. Statistical significance was addressed by two-tailed Student’s t test. * P ≤ 0.05; *** P ≤ 0.001; **** P ≤ 0.0001. Figure elements were created with BioRender.

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Immunofluorescence, Staining, Fluorescence, Two Tailed Test

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) Hyperpigmentation in palmar and plantar aspects of the fore- and hindpaws of Ten1 hom mice at P23 ( n WT/hom: 5/9). ( B ) H&E staining of Ten1 hom snout (P5) and flank (P23) skin. Melanin deposition demonstrated by Fontana-Masson staining (see arrows). Five days ( n WT/hom: 2/4) and 23 days ( n WT/hom: 6/6). ( C ) Tongue H&E staining at P23 ( n WT/hom: 6/7) and P32 ( n WT/hom: 1/1) showed hyperkeratosis, decreased number of papillae, architectural disorganization, and nuclear pleomorphism in Ten1 hom mice. ( D ) Decreased villi length and signs of mucosal atrophy in the small intestine of Ten1 hom mice at P23 ( n WT/hom: 6/6).

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Staining

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) H&E-stained femoral bone marrow at P5 and P23 of control and hom mice (P5 n WT/hom: 2/3; P23 n WT/hom: 3/3). ( B ) Thymic atrophy in P23 Ten1 hom animals (P5 n WT/hom: 2/7; P23 n WT/hom: 6/7).

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Staining, Control

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) Representative pictures of H&E-stained cerebellum sections at P0.5 ( n WT/hom: 6/6), P5 ( n WT/hom: 5/8), and P23 ( n WT/hom: 6/5) at the same magnification. ( B ) Double immunofluorescence of granular (asterisk; NeuN, red) and Purkinje (arrow; calbindin, green) cells in the cerebellum of Ten1 hom animals at P23 ( n WT/hom: 4/4). ( C ) Ocular changes in P21 Ten1 hom animals: Anterior synechia and retinal alterations at P21 are displayed ( n WT/hom: 3/5).

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Staining, Immunofluorescence

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: Comparison of human TBD clinical manifestations and Ten1 knockout mouse phenotypes. Reported frequencies in NCI DC/TBD cohort modified from ( 10 ). Symptoms with less than 5% frequency were omitted. Hoyeraal-Hreidarsson syndrome modified from ( 50 ). KO, knockout; n.d., not detected; NA, not analyzed; BMD, bone mineral density.

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Comparison, Knock-Out, Modification

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) Mki67 gene expression levels for cell proliferation analysis in the cerebrum, cerebellum, liver, lung, and skin at P23. * P ≤ 0.05; **** P ≤ 0.0001. ( B ) IHC showed fewer Ki67-positive cells in Ten1 hom mice compared to controls in cerebellum at P5 and in skin and small intestine at P23. ( C ) TUNEL staining for apoptosis in Ten1 hom animals in cerebellum at P5 and in skin and small intestine at P23. Figure elements in (A) were created with BioRender. For (B) and (C): n WT/hom: P5 2/4; P23 6/7.

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Gene Expression, TUNEL Assay, Staining

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: Representative pictures of p53 ( A ) and p21 ( B ) IHC in the cerebellum, small intestine, and spleen from control and Ten1 hom mice at P5 (left) and P23 (right). The area inside the rectangles of the low magnification pictures on top is shown below on a higher magnification. For (A) and (B): n WT/hom: P5 2/4; P23 6/7.

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Control

Journal: Life Science Alliance

Article Title: LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

doi: 10.26508/lsa.202101050

Figure Lengend Snippet: (A) Primary, wild type rat astrocytes were fixed with 3% PFA and stained for cilia using mouse anti-Arl13b (green) and rabbit anti-RILPL2 (red). Box indicates the region enlarged in the image shown at right. Bar, 10 μm. Arrows indicate the base of the cilium. (B) hTERT-RPE cells were transfected with HA-RILPL2 at 80% confluency and 24 h later either medium changed (+FBS) or serum starved (−FBS) for 2 h. Cells were fixed with −20°C methanol for 2 min and stained with rabbit or mouse anti-HA to detect RILPL2 (red) and mouse anti-CEP164, or rabbit anti-CP110 (green). Arrows indicate the location of the centriolar region. Bar, 2.5 μm. (C) Quantitation of percent of cells that show centriolar localized RILPL2 ± 2 h serum starvation. Significance was determined by t test; * P = 0.03. Error bars represent standard error of the mean from two independent experiments with >50 cells per condition. (D) hTERT-RPE cells expressing mKO2-PACT (magenta) were transfected with GFP-RILPL2 (green). Cells were serum starved and 15 min later imaged by capturing Z-stacks every 6 min for the next 8 h. Yellow arrowheads indicate the location of the centrosome as marked by mKO2-PACT. Upper panel shows GFP-RILPL2 in grayscale. Scale bar, 5 μm. (E) Times at which GFP-RILPL2 appeared at the mKO2-PACT structure. Error bar represents standard error of the mean.

Article Snippet: RILPL2 knockout mouse line was originally acquired from The KOMP Repository and is now available from MMRRC.org (Stock number: 049453-UCD).

Techniques: Staining, Transfection, Quantitation Assay, Expressing

Journal: Life Science Alliance

Article Title: LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

doi: 10.26508/lsa.202101050

Figure Lengend Snippet: (A) RPE cells transfected with HA-RILP2 or GFP-RILPL2 as indicated and stained for endogenous MyoVa. Bars, 10 μm. (B) R1441C MEFs transfected with HA-RILPL2, ± MLi-2 LRRK2 inhibitor. Cells were serum starved for 16 h before fixation and staining with mouse anti-CEP164 or rabbit anti-HA. Bars, 10 μm. Boxes indicate the areas enlarged at the right. (C) Quantitation of pericentriolar fluorescence intensity in arbitrary units ±MLi-2. * P = 0.024. (D) HEK293T cells depleted of MyoVa as indicated and transfected with GFP-R1441G LRRK2 for 24 h. Cells were stained with rabbit anti-pRab10 antibody. (D, E) Quantitation of percent of cells with perinuclear, clustered Rab10 in control or MyoVa shRNA treated cells as in (D).

Article Snippet: RILPL2 knockout mouse line was originally acquired from The KOMP Repository and is now available from MMRRC.org (Stock number: 049453-UCD).

Techniques: Transfection, Staining, Quantitation Assay, Fluorescence, shRNA

Journal: Life Science Alliance

Article Title: LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

doi: 10.26508/lsa.202101050

Figure Lengend Snippet: (A) Localization of pRab10 and the centriole marker, Centrin 3. HeLa cells were transfected with R1441G Myc-LRRK2 and methanol fixed for staining with rabbit anti-pRab10 and mouse anti-Centrin 3. The boxed area is enlarged in the adjacent panels at right. Bar, 10 μM. (B) HEK293T cells were transfected with R1441G LRRK2, full-length MyoVa mCherry, and HA-RILPL2 for 16 h, treated ± 200 nM MLi-2 for 2 h and stained with mouse anti-HA and rabbit anti-phosphoRab10. Bar, 10 μm. (C) Quantitation of cells with perinuclear RILPL2 from the experiment in (B) from two independent experiments with >50 cells per condition. *0.027; **0.0026 t test. (B, D) HEK293T cells depleted of MyoVa were transfected with R1441G LRRK2 and HA-RILPL2 and stained for RILPL2 and pRab10 as in (B). (E) Immunoblot of HEK293T cells depleted of MyoVa using three shRNAs to create stable cell lines. Numbers at left in this and subsequent figures represent mass in kilodaltons. (F) Quantitation of perinuclear RILPL2 in cells with and without MyoVa. No significant difference was detected.

Article Snippet: RILPL2 knockout mouse line was originally acquired from The KOMP Repository and is now available from MMRRC.org (Stock number: 049453-UCD).

Techniques: Marker, Transfection, Staining, Quantitation Assay, Western Blot, Stable Transfection

Journal: Life Science Alliance

Article Title: LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

doi: 10.26508/lsa.202101050

Figure Lengend Snippet: (A) HEK293T cells were co-transfected with Myc-LRRK2-R1441G and MyoVa-mCherry. After 18 h, cells were fixed and stained with mouse anti-Myc (blue) and rabbit anti-pRab10 (cyan). (B) Quantitation of colocalization by Mander’s method between MyoVa and pRab10. The fraction of MyoVa that colocalizes with pRab10 and vice-versa are quantified. Error bars indicate standard error of the mean of colocalization coefficient for >20 cells. (A, C) HEK293T cells depleted of Rab10 (KD) were co-transfected with Myc-LRRK2 R1441G and HA-RILPL2. (A) After 18 h, the cells were stained as in (A). (D) R1441G LRRK2 MEF cells, either control or depleted of RILPL2, transfected with globular tail domain-mCherry. Markers are indicated; cells were stained wisth mouse anti-CEP164 or rabbit anti-mCherry. (E) Quantitation of the experiment shown in (D); P -values were determined by t test: ***0.0009 and 0.0004. (F) qPCR quantitation of RILPL2 mRNA in cells treated with two different shRNAs as indicated.

Article Snippet: RILPL2 knockout mouse line was originally acquired from The KOMP Repository and is now available from MMRRC.org (Stock number: 049453-UCD).

Techniques: Transfection, Staining, Quantitation Assay

Journal: Life Science Alliance

Article Title: LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

doi: 10.26508/lsa.202101050

Figure Lengend Snippet: (A) MyoVa or MyoVa ΔD-mCherry and GFP-LRRK2-R1441G were co-transfected into HEK293T cells; 24 h post transfection, cells were incubated with 200 nM MLi-2 for 4 h. Cells were lysed in lysis buffer and 400 μg extract immunoprecipitated with anti-RFP antibodies on protein G beads. Samples (50% of IP and 15% of input for all IPs in all panels) were immunoblotted using anti-GFP, anti-RFP, anti-tubulin, anti-Rab10 and anti-pRab10 antibodies. (A, B) Quantitation of the relative amount of Rab10 (left) or pRab10 (right) co-immunoprecipitated by MyoVa and MyoVa ΔD in (A). Error bars indicate standard error of the mean from two gels per co-IP. Significance was determined by t test; *** P = 0.0003; ns, not significant with P = 0.295. (C) MyoVa globular tail domain (GTD)-mCherry and GFP-LRRK2 R1441G were co-transfected into HEK293T cells. After 24 h, cells were incubated ± 200 nM MLi-2 for 4 h. Cells were lysed in lysis buffer and immunoprecipitated with anti-RFP antibodies on protein G beads. Samples were immunoblotted with anti-GFP, anti-RFP, anti-Rab10 and anti-pRab10 antibodies. (D) RILPL2-GFP, MyoVa GTD-mCherry, and Myc-LRRK2 R1441G were co-transfected into HEK293T cells; 24 h post-transfection, cells were incubated ± 200 nM MLi-2 for 4 h. Extracts in lysis buffer were immunoprecipitated with GFP-binding protein Sepharose. Samples were immunoblotted with anti-GFP, anti-RFP, anti-Rab10 and anti-pRab10 antibodies. (D, E) Quantitation of the relative amount of total MyoVa GTD (left) and Rab10 (right) co-immunoprecipitated by RILPL2-GFP under the conditions indicated below (E). Significance was determined by t test; * P = 0.021 and 0.035 (left) and P = 0.014 and 0.024 (right).

Article Snippet: RILPL2 knockout mouse line was originally acquired from The KOMP Repository and is now available from MMRRC.org (Stock number: 049453-UCD).

Techniques: Transfection, Incubation, Lysis, Immunoprecipitation, Quantitation Assay, Co-Immunoprecipitation Assay, Binding Assay

Journal: Life Science Alliance

Article Title: LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

doi: 10.26508/lsa.202101050

Figure Lengend Snippet: (A) hTERT-RPE cells were transfected with HA-RILPL2 or mock transfected with lipofectamine and 24 h later, serum-starved overnight to initiate ciliogenesis. Cells were fixed with ice cold −20°C methanol for 2 min and co-stained with rabbit anti-HA (green) or mouse anti-Arl13b (red). Arrowhead indicates the likely location of a RILPL2-positive centriole. Dotted lines indicate cell outlines. Bar, 10 μm. (B) Quantitation of cells with cilia ± HA-RILPL2 transfection. Error bars represent SEM from two experiments, each with >25 cells per condition. Significance was determined by t test; * P = 0.0164. (C) WT, RILPL1, and RILPL2 knockout A549 cells were plated at 80% confluency. After 24 h, cells were serum starved by transfer into 2% FBS-containing medium for 48 h. Cells were fixed and stained with mouse anti-Arl13b antibody for quantitation of cells with cilia. Error bars represent SEM from two experiments with >100 cells per condition in each experiment. #1 and #2 indicate two different A549 RILPL2 knockout clonal cell lines. (D) Quantitation of cells with two CP110 dots indicating capped centrioles in: (left) hTERT-RPE cells ± HA-RILPL2 for 24 h; (right) hTERT-RPE cells ± MyoVa knock-down. Cells were serum starved for 2 h, fixed with −20°C methanol for 2 min and co-stained with anti-HA, anti-CP110 or anti-CEP164/CP110 antibodies. Error bars represent SEM from two experiments with >25 cells per condition in each experiment. Significance was determined by t test; ** P = 0.0067 and 0.0045. (B, E) RPE cells expressing HA-RILPL2 as in panel (B) were scored for TTBK2 at the centriole after serum starvation as in . Shown are data from two independent experiments; >25 cells per experiment. No significant difference was detected. (B, F) RPE cells expressing HA-RILPL2 as in panel (B) were scored for endogenous MyoVa at the centriole; shown are data from two independent experiments; >50 cells per experiment for each condition. **** P < 0.0001 by t test.

Article Snippet: RILPL2 knockout mouse line was originally acquired from The KOMP Repository and is now available from MMRRC.org (Stock number: 049453-UCD).

Techniques: Transfection, Staining, Quantitation Assay, Knock-Out, Expressing

Journal: Life Science Alliance

Article Title: LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

doi: 10.26508/lsa.202101050

Figure Lengend Snippet: (A) Top row, RPE cells transfected with HA-RILPL2 (red) and stained for rabbit anti-CP110 (green). Cells were serum-starved for 2 h before fixation and staining. Inset 1, RILPL2 transfected cell; inset 2 is a non-RILPL2 transfected cell. Second row, MyoVa-depleted RPE cells; shown is CP110 and CEP164 staining, with the boxed area enlarged at right. (B) TTBK2 and centriole marker, rootletin localization in RPE cells expressing HA-RILPL2 or HA alone as indicated. Boxed regions in the first column are enlarged at right. (C) As in (B), monitoring MyoVa and rootletin localization. Shown are two cells, with and without RILPL2 expression.

Article Snippet: RILPL2 knockout mouse line was originally acquired from The KOMP Repository and is now available from MMRRC.org (Stock number: 049453-UCD).

Techniques: Transfection, Staining, Marker, Expressing

Journal: The Plant Cell

Article Title: The WRKY6 Transcription Factor Modulates PHOSPHATE1 Expression in Response to Low Pi Stress in Arabidopsis [W] [OA]

doi: 10.1105/tpc.108.064980

Figure Lengend Snippet: Repression of PHO1 Expression by WRKY6 Was Released in Response to Low Pi Stress.

Article Snippet: We thank Imre E. Somssich (Max-Planck-Institut für Züchtungsforschung, Abteilung Biochemie, Germany) for kindly providing At WRKY6 overexpression lines 35S: WRKY6-3 , 35S: WRKY6-5 , and 35S: WRKY6-9 and the At WRKY6 knockout mutant ( wrky6-1 ).

Techniques: Expressing

Journal: The Plant Cell

Article Title: The WRKY6 Transcription Factor Modulates PHOSPHATE1 Expression in Response to Low Pi Stress in Arabidopsis [W] [OA]

doi: 10.1105/tpc.108.064980

Figure Lengend Snippet: ChIP Assays for At WRKY6 Binding to the W-Box of the PHO1 Promoter in Vivo.

Article Snippet: We thank Imre E. Somssich (Max-Planck-Institut für Züchtungsforschung, Abteilung Biochemie, Germany) for kindly providing At WRKY6 overexpression lines 35S: WRKY6-3 , 35S: WRKY6-5 , and 35S: WRKY6-9 and the At WRKY6 knockout mutant ( wrky6-1 ).

Techniques: Binding Assay, In Vivo

Journal: The Plant Cell

Article Title: The WRKY6 Transcription Factor Modulates PHOSPHATE1 Expression in Response to Low Pi Stress in Arabidopsis [W] [OA]

doi: 10.1105/tpc.108.064980

Figure Lengend Snippet: Suppression of PHO1 Expression by WRKY6.

Article Snippet: We thank Imre E. Somssich (Max-Planck-Institut für Züchtungsforschung, Abteilung Biochemie, Germany) for kindly providing At WRKY6 overexpression lines 35S: WRKY6-3 , 35S: WRKY6-5 , and 35S: WRKY6-9 and the At WRKY6 knockout mutant ( wrky6-1 ).

Techniques: Expressing

Journal: The Plant Cell

Article Title: The WRKY6 Transcription Factor Modulates PHOSPHATE1 Expression in Response to Low Pi Stress in Arabidopsis [W] [OA]

doi: 10.1105/tpc.108.064980

Figure Lengend Snippet: ChIP-qPCR Assays to Detect the Association of WRKY6 and the PHO1 Promoter in the Tested Plants as Indicated under Pi-Sufficient (MS) and Pi-Deficient (LP) Conditions.

Article Snippet: We thank Imre E. Somssich (Max-Planck-Institut für Züchtungsforschung, Abteilung Biochemie, Germany) for kindly providing At WRKY6 overexpression lines 35S: WRKY6-3 , 35S: WRKY6-5 , and 35S: WRKY6-9 and the At WRKY6 knockout mutant ( wrky6-1 ).

Techniques: ChIP-qPCR

Journal: The Plant Cell

Article Title: The WRKY6 Transcription Factor Modulates PHOSPHATE1 Expression in Response to Low Pi Stress in Arabidopsis [W] [OA]

doi: 10.1105/tpc.108.064980

Figure Lengend Snippet: WRKY6 Protein Blot Analysis.

Article Snippet: We thank Imre E. Somssich (Max-Planck-Institut für Züchtungsforschung, Abteilung Biochemie, Germany) for kindly providing At WRKY6 overexpression lines 35S: WRKY6-3 , 35S: WRKY6-5 , and 35S: WRKY6-9 and the At WRKY6 knockout mutant ( wrky6-1 ).

Techniques:

Journal: Molecular microbiology

Article Title: Bactericidal peptidoglycan recognition protein induces oxidative stress in Escherichia coli through a block in respiratory chain and increase in central carbon catabolism

doi: 10.1111/mmi.13733

Figure Lengend Snippet: (A) Mutants with increased resistance to PGRP killing were identified by three-stage screening of the entire Keio collection of single gene deletion mutants (Fig. S1 and Table S1) and here the survival of the parental strain (BW25113) and mutants following 3-hr incubation with 200 µg/ml of BSA (as a control) or PGRP is shown. Gene products, their functions, and numerical data are shown in Table S1. (B) Mutants for the key genes for the respiratory chain and TCA cycle, and their regulators (cyaA and crp) were constructed in MG1655 and their sensitivity to killing by 100 µg/ml of PGRP was similarly tested. The results are means of 3 experiments, expressed as percent of initial inoculum (100%) + SEM; ^ P≤0.05, ^^ P<0.001, numbers of viable bacteria (colony forming units) of Δ mutants versus parental strain (t-test).

Article Snippet: Bacteria, growth, and media The entire Keio collection of 3884 E. coli K-12 in-frame single-gene knockout mutants was obtained from the National BioResource Project, National Institute of Genetics, Japan ( Baba et al ., 2006 ).

Techniques: Incubation, Control, Construct, Bacteria

Journal: Molecular microbiology

Article Title: Bactericidal peptidoglycan recognition protein induces oxidative stress in Escherichia coli through a block in respiratory chain and increase in central carbon catabolism

doi: 10.1111/mmi.13733

Figure Lengend Snippet: (A) Time kinetics of changes of H2O2 in E. coli MG1655 treated with 100 µg/ml BSA or PGRP, or with 100 µM paraquat. (B) H2O2 in E. coli BW25113 and in the indicated deletion mutants from Keio collection treated with 100 µg/ml BSA or PGRP for 15 min. (C) H2O2 in E. coli MG1655 and in the indicated deletion mutants constructed in our laboratory treated with 100 µg/ml BSA or PGRP for 15 min. The results are means of 3–4 experiments ± SEM (SEM were within symbols if not visible); * P<0.05, ** P<0.001, PGRP vs BSA; +P<0.05, ++P<0.001, paraquat vs BSA; ^ P<0.05, ^^ P<0.001, PGRP-treated mutant vs parental strain.

Article Snippet: Bacteria, growth, and media The entire Keio collection of 3884 E. coli K-12 in-frame single-gene knockout mutants was obtained from the National BioResource Project, National Institute of Genetics, Japan ( Baba et al ., 2006 ).

Techniques: Construct, Mutagenesis

Journal: Molecular microbiology

Article Title: Bactericidal peptidoglycan recognition protein induces oxidative stress in Escherichia coli through a block in respiratory chain and increase in central carbon catabolism

doi: 10.1111/mmi.13733

Figure Lengend Snippet: Parental E. coli or the indicated deletion mutants from Keio collection (BW25113) or from our laboratory (MG1655) were treated with 100 µg/ml BSA or PGRP for 15 min (A) or 5 min (B and C). The results are means of 3–4 experiments + SEM; * P<0.05, ** P<0.001, PGRP vs BSA; ^ P<0.05, ^^ P<0.001, mutant vs parental strain.

Article Snippet: Bacteria, growth, and media The entire Keio collection of 3884 E. coli K-12 in-frame single-gene knockout mutants was obtained from the National BioResource Project, National Institute of Genetics, Japan ( Baba et al ., 2006 ).

Techniques: Mutagenesis

Journal: Molecular microbiology

Article Title: Bactericidal peptidoglycan recognition protein induces oxidative stress in Escherichia coli through a block in respiratory chain and increase in central carbon catabolism

doi: 10.1111/mmi.13733

Figure Lengend Snippet: E. coli and the indicated deletion mutants from Keio collection (BW25113) or from our laboratory (MG1655) were treated with 100 µg/ml BSA or PGRP for 30 min. The results are means of 3 experiments + SEM; * P<0.05, ** P<0.001, PGRP vs BSA; ^ P<0.05, mutant vs parental strain.

Article Snippet: Bacteria, growth, and media The entire Keio collection of 3884 E. coli K-12 in-frame single-gene knockout mutants was obtained from the National BioResource Project, National Institute of Genetics, Japan ( Baba et al ., 2006 ).

Techniques: Mutagenesis

Journal: The Plant Cell

Article Title: Ethylene Responses in Rice Roots and Coleoptiles Are Differentially Regulated by a Carotenoid Isomerase-Mediated Abscisic Acid Pathway ^[OPEN]

doi: 10.1105/tpc.15.00080

Figure Lengend Snippet: Genetic Interactions between mhz5-1 and Ethylene Receptor Loss-of-Function Mutants through Double Mutant Analyses. (A) Comparison of the root ethylene response in Nipponbare (Nip), Dongjin (DJ), and the single and double mutants in the absence or presence of ethylene (1 ppm). Representative 2.5-d-old dark-grown seedlings are shown. Bars = 10 mm. (B) Ethylene dose–response curves for the root length of 2.5-d-old dark-grown seedlings of Nipponbare, Dongjin, mhz5-1, and double mutants (ers1 mhz5-1, ers2 mhz5-1, and etr2 mhz5-1). The values are the means ± sd of 20 to 30 seedlings per genotype at each dose. The experiment was repeated at least three times with similar results.

Article Snippet: The T-DNA knockout mutants ers1 , ers2 , and etr2 are in the DJ background and were obtained from the POSTECH Biotech Center ( Yi and An, 2013 ).

Techniques: Mutagenesis, Comparison