Journal: Scientific Reports

Article Title: Modulation of the p75 neurotrophin receptor suppresses age-related basal forebrain cholinergic neuron degeneration

doi: 10.1038/s41598-019-41654-8

Figure Lengend Snippet: Diminished efficacy with shorter and later treatment. Five weeks of treatment once daily with 50 mg/kg LM11A-31 (n = 5) beginning at 17 months of age partially prevented and/or reversed the decrease in cholinergic neuron area in the MS and VDB seen in the vehicle treated mice (n = 4). Cholinergic neurons were identified by ChAT immunoreactivity and an unbiased estimate of area was obtained using Microbrightfield (MBF) software. A significant decrease in cell area measurements was seen in all groups (t-test, (n = 8) vs 18 months (n = 4), p values = 0.018–0.001). Inhibition and/or reversal of loss of cell size after five weeks of treatment was partial and reached significance only in the composite data for MS + VDB (t-test, treated vs vehicle, p = 0.032). Values are expressed as mean + sem.

Article Snippet: Sections were coded to reduce bias and Microbrightfield (MBF) unbiased stereology was used for the collection of cellular data.

Techniques: Software, Inhibition

Journal: The New Phytologist

Article Title: The Medicago truncatula Vacuolar iron Transporter‐Like proteins VTL4 and VTL8 deliver iron to symbiotic bacteria at different stages of the infection process

doi: 10.1111/nph.16735

Figure Lengend Snippet: Development of an iron reporter for symbiotic rhizobium bacteria. (a) Diagram of the bacterial P mbfA:lux reporter and regulation of its expression by the iron response regulator (Irr) which binds the upstream Iron Control Element (ICE). In iron‐replete conditions, Irr binds iron in the form of haem and is degraded, leading to de‐repression (activation) of target gene expression. (b) Expression of the endogenous mbfA gene in free‐living Sinorhizobium meliloti 1021 in response to iron. Bacteria were grown in UMS medium without (−Fe) or with 40 µM iron sulphate (+Fe) until mid‐log phase, then harvested for RNA extraction. mbfA transcript levels were assayed by RT‐qPCR. Values were normalized to the housekeeping gene gapA, and are the mean ± SE of three biological replicates ( ***, P = 0.0001, Bio‐Rad CFX Maestro software). (c) Sinorhizobium meliloti 1021 carrying the lux plasmid without promoter (P‐: lux ); the P mbfA:lux reporter; P mbfA:lux with a mutated ICE motif; or a constitutive lux reporter with the nptII promoter were grown as in (b). Luminescence was measured in a plate reader and corrected for cell density (OD 600 ). Values are the mean ± SE of three biological replicates (cell cultures grown in parallel). A.U., arbitrary units. (d) P mbfA:lux activity normalized for constitutive lux activity of the P nptII:lux reporter in response to iron. Error bars represent SE.

Article Snippet: Plants growing on Terragreen–sand mixture were inoculated with S. meliloti 1021 carrying the P mbfA:lux reporter plasmid or the mutated P mbfA ICE :lux reporter at 7 d post‐germination, and grown for a further 21 d. Plants were dug up, roots rinsed with distilled water, blotted dry and imaged using the NightOWL II LB 983 in vivo imaging system with I ndi GO software (Berthold Technologies, Bad Wildbad, Germany).

Techniques: Bacteria, Expressing, Control, Activation Assay, Targeted Gene Expression, RNA Extraction, Quantitative RT-PCR, Software, Plasmid Preparation, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Genome-wide Characterization of the MBF1 Gene Family and Its Expression Pattern in Different Tissues and Under Stresses in Medicago truncatula and Medicago sativa

doi: 10.3390/ijms26020455

Figure Lengend Snippet: Properties of the predicted MBF1 proteins in M. truncatula and M. sativa .

Article Snippet: By investigating the tissue-specific expression patterns of all the MBF1 genes in Medicago , we found that MBF1 was mainly expressed in the leaves of these plants.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Genome-wide Characterization of the MBF1 Gene Family and Its Expression Pattern in Different Tissues and Under Stresses in Medicago truncatula and Medicago sativa

doi: 10.3390/ijms26020455

Figure Lengend Snippet: Phylogenetic relationships, motifs, and gene structures of MBF1 genes. ( A ) Phylogenetic analysis of MBF1 families across Medicago , Arabidopsis , Oryza sativa , Zea mays , Vigna unguiculata , Brassica rapa , algae, bryophytes, pteridophytes, and gymnosperms. Full-length protein sequences of MBF1s were constructed using MEGA-X based on the neighbor-joining (NJ) method with 1000 bootstraps. Blue triangles, red circles, green squares, and orange pentagrams represent algae, bryophytes, pteridophytes, and gymnosperms, respectively. ( B ) Phylogenetic relationships, motifs, and gene structures of the MBF1 genes from M. truncatula and M. sativa . The motifs are indicated by different colored boxes with different numbers. Blue boxes indicate 5′- and 3′-untranslated regions; green boxes indicate exons; black lines indicate introns.

Article Snippet: By investigating the tissue-specific expression patterns of all the MBF1 genes in Medicago , we found that MBF1 was mainly expressed in the leaves of these plants.

Techniques: Algae, Construct

Journal: International Journal of Molecular Sciences

Article Title: Genome-wide Characterization of the MBF1 Gene Family and Its Expression Pattern in Different Tissues and Under Stresses in Medicago truncatula and Medicago sativa

doi: 10.3390/ijms26020455

Figure Lengend Snippet: Chromosome distributions of MBF1s in M. truncatula and M. sativa . The chromosomal location and interchromosomal relationship of M. truncatula ( A ) and M. sativa ( B ). ( C ) Synteny analysis of MBF1 genes between Arabidopsis , M. truncatula , and M. sativa . The gray lines in the background indicate collinear blocks between M. truncatula , and M. sativa / Arabidopsis , while the blue lines highlight syntenic MBF1 gene pairs. ( D ) The Ka/Ks values of MBF1 gene pairs for Mt-Ms .

Article Snippet: By investigating the tissue-specific expression patterns of all the MBF1 genes in Medicago , we found that MBF1 was mainly expressed in the leaves of these plants.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Genome-wide Characterization of the MBF1 Gene Family and Its Expression Pattern in Different Tissues and Under Stresses in Medicago truncatula and Medicago sativa

doi: 10.3390/ijms26020455

Figure Lengend Snippet: Putative cis -elements and transcription factor binding sites in the promoter regions of MBF1 genes from M. truncatula and M. sativa . ( A ) The color and number of the grid indicate the numbers of different cis -acting elements in these MBF1 genes. ( B ) The colored block represent different types of cis -acting elements and their locations in each MBF1 gene.

Article Snippet: By investigating the tissue-specific expression patterns of all the MBF1 genes in Medicago , we found that MBF1 was mainly expressed in the leaves of these plants.

Techniques: Binding Assay, Blocking Assay

Journal: International Journal of Molecular Sciences

Article Title: Genome-wide Characterization of the MBF1 Gene Family and Its Expression Pattern in Different Tissues and Under Stresses in Medicago truncatula and Medicago sativa

doi: 10.3390/ijms26020455

Figure Lengend Snippet: Subcellular localization assay of MBF1s. The coding region of MBF1 s was cloned into the pCAMBIA1300-35S-eGFP vector. This construct was expressed transiently in Arabidopsis mesophyll protoplasts using the PEG-mediated method. eGFP, enhanced GFP. Scale bar = 10 μm.

Article Snippet: By investigating the tissue-specific expression patterns of all the MBF1 genes in Medicago , we found that MBF1 was mainly expressed in the leaves of these plants.

Techniques: Clone Assay, Plasmid Preparation, Construct

Journal: International Journal of Molecular Sciences

Article Title: Genome-wide Characterization of the MBF1 Gene Family and Its Expression Pattern in Different Tissues and Under Stresses in Medicago truncatula and Medicago sativa

doi: 10.3390/ijms26020455

Figure Lengend Snippet: Determining MBF1 expression patterns in different tissue sites by qRT-PCR. ( A ) Expression levels of MtMBF1 genes in various tissues verified by qRT-PCR. ( B ) Expression levels of MsMBF1 genes in various tissues verified by qRT-PCR.

Article Snippet: By investigating the tissue-specific expression patterns of all the MBF1 genes in Medicago , we found that MBF1 was mainly expressed in the leaves of these plants.

Techniques: Expressing, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Genome-wide Characterization of the MBF1 Gene Family and Its Expression Pattern in Different Tissues and Under Stresses in Medicago truncatula and Medicago sativa

doi: 10.3390/ijms26020455

Figure Lengend Snippet: Quantification of the expression levels of the selected MBF1 genes from M. truncatula and M. sativa under abiotic stress. Quantification was conducted using qRT-PCR at 0 h, 1 h, 3 h, 6 h, 12 h, 24 h, and 48 h after NaCl, PEG, heat, and cold stress treatments. Expression levels were normalized to the reference gene ACTIN (JQ028730), and RNA was extracted from whole seedlings. Data are the average of three independent biological samples ±SE; the vertical bars indicate standard deviations. ** indicates p < 0.01, and * indicates p < 0.05. Pearson correlation coefficients (“Pearson”) for expression levels under NaCl, PEG, cold, and heat stresses are shown.

Article Snippet: By investigating the tissue-specific expression patterns of all the MBF1 genes in Medicago , we found that MBF1 was mainly expressed in the leaves of these plants.

Techniques: Expressing, Quantitative RT-PCR