Journal: PLoS Pathogens

Article Title: The TFPI-2 Derived Peptide EDC34 Improves Outcome of Gram-Negative Sepsis

doi: 10.1371/journal.ppat.1003803

Figure Lengend Snippet: ( A ) E. coli ATCC 25922 and P. aeruginosa PA01 bacteria were incubated 30–60 min with citrate plasma alone (Control) or supplemented with EDC34 (at 3 µM) at 37°C. The bacterial cells and supernatants were collected and proteins were detected by immunoblotting with antibodies recognizing C1q or C5b-9. CP, citrate plasma; S, supernatant or unbound bacteria; P, pellet or material bound to bacterial cells. ( B ), as in , but antibodies against C3a were used. ( C ) Quantification of deposition of complement components on bacteria by flow cytometry, left panel , comparison of the mean proportion of bacteria positive for C1q/C3a binding in citrate plasma and in plasma supplemented with EDC34. Right panel , comparative degree of C1q and C3a binding to E. coli and P. aeruginosa strains, expressed as means of the fluorescence index ( FI ; proportion of bacteria positive for C1q/C3a multiplied by the mean intensity of C1q/C3a binding). (n = 4, mean±SEM is presented; Two-Way ANOVA Bonferroni's Multiple Comparison Test). ( D ) TFPI-2 and C3a binding to bacteria in fibrin slough collected from an infected chronic wound were visualized by using gold-labeled antibodies of different sizes, specific for the C-terminus of TFPI-2 (20 nm) and C3a (10 nm), respectively. Insert shows a higher magnification.

Article Snippet: For immunostaining , rabbit polyclonal antibodies against the C-terminal of TFPI-2 (CAKALKKKKKMPKLRFASRIRKIRKKQF) alone, or in combination with rabbit polyclonal antibodies against the C-terminal part of C3a (LGE27 antibodies) (1 μg/ml) (Innovagen AB) were utilized.

Techniques: Bacteria, Incubation, Clinical Proteomics, Control, Western Blot, Flow Cytometry, Comparison, Binding Assay, Fluorescence, Infection, Labeling

Journal:

Article Title: ? N -p73? Accumulates in Human Neuroblastic Tumors

doi:

Figure Lengend Snippet: Genomic organization of the p73 gene. Diagram showing the various domains studied by RT-PCR, in orange boxes spanning: TAD, transactivating domain; DBD, DNA specific binding domain; OD, oligomerization domain. Exons are represented by boxes of various colors: N-terminal part: green for normal exons; yellow for alternate 1-bis and 3-bis; core domain, gray; C-terminal part, blue. P1, P2 and P3 indicate the three promoters of the gene. The size of amplicons is indicated in brackets. The thin lines represent introns.

Article Snippet: To detect p73 localization, the primary antibody (Ab) was a rabbit polyclonal Ab raised against p73 human (provided by Sanofi-Recherche, Labège) at a dilution of 1/300 in phosphate-buffered saline (PBS).

Techniques: Reverse Transcription Polymerase Chain Reaction, Binding Assay

Journal:

Article Title: ? N -p73? Accumulates in Human Neuroblastic Tumors

doi:

Figure Lengend Snippet: p73α immunocytochemistry in SH-SY5Y cells. A–C: Control SH-SY5Y cells transfected by empty plasmid. Note a nuclear p73α immunostaining in (C) compared to the negative control without p73 antibody in (B). D–F: FGF1 overexpression mediates neuritic outgrowth in a FGF1-transfected clone (D); negative control (E). Arrows indicate punctuate cytoplasmic p73α dotted staining in differentiated cells (F). G–I: SH-SY5Y cells treated by exogenous NGF + aphidicolin for 8 days. Note the strong p73α cytoplasmic staining which contrasts with the light nuclear staining; H: negative control. B, E, H: magnification, ×200; A, C, D, F, G, I: magnification, ×400.

Article Snippet: To detect p73 localization, the primary antibody (Ab) was a rabbit polyclonal Ab raised against p73 human (provided by Sanofi-Recherche, Labège) at a dilution of 1/300 in phosphate-buffered saline (PBS).

Techniques: Immunocytochemistry, Control, Transfection, Plasmid Preparation, Immunostaining, Negative Control, Over Expression, Staining

Journal:

Article Title: ? N -p73? Accumulates in Human Neuroblastic Tumors

doi:

Figure Lengend Snippet: RT-PCR analysis showing various transcript species spanning the three domains of the p73 gene. GNB: ganglioneuroblastoma; NB, undifferentiated neuroblastoma. A: N-terminal part: P1 transcripts (lanes 2, 4, and 6, P3 transcripts (lanes 8, 9, and 10). Note that transcript 1bis-2–3 (initiated from P2) is barely detectable with no significant differences in intensity between NB and GNB according to semiquantitative RT-PCR normalized by GAPDH gene. A faint upper 1-2-3 band (lanes 2 and 6) is considered as non-specific PCR product. Occasionally, a transcript species arising from another promoter denoted as P2 (Figure 1) ▶ was expressed with faint intensity (lanes 1, 3, and 5). In lanes 2 and 6, a Δ-exon 2 band in a NB/GNB and a NB specimen is barely detectable (a ratio r<0.5, is considered 0 according to the scale of Table 1 ▶ ). B: Core and C-terminal transcripts. Exons 6–8 transcript (lanes 1, 2, and 3). Exons 12–14 transcripts (lanes 5, 6, and 7). M, size marker. C: To illustrate the differential P1 usage in lymphocytes versus NTs, RT-PCR products separated in 4% polyacrylamide gel showed P1-Δexon 2 transcript which was consistently found in various histology (NB, NB/GNB, GNB, GNB/GN) whereas it was absent or expressed at barely detectable levels in corresponding lymphocytes.

Article Snippet: To detect p73 localization, the primary antibody (Ab) was a rabbit polyclonal Ab raised against p73 human (provided by Sanofi-Recherche, Labège) at a dilution of 1/300 in phosphate-buffered saline (PBS).

Techniques: Reverse Transcription Polymerase Chain Reaction, Marker

Journal:

Article Title: ? N -p73? Accumulates in Human Neuroblastic Tumors

doi:

Figure Lengend Snippet: Fold Increases of p73 Transcript Species

Article Snippet: To detect p73 localization, the primary antibody (Ab) was a rabbit polyclonal Ab raised against p73 human (provided by Sanofi-Recherche, Labège) at a dilution of 1/300 in phosphate-buffered saline (PBS).

Techniques:

Journal: Cell Reports

Article Title: Vesicular Glutamate Transporters (SLCA17 A6, 7, 8) Control Synaptic Phosphate Levels

doi: 10.1016/j.celrep.2020.108623

Figure Lengend Snippet: VGLUT2 Is a Key Factor of Synaptic [Pi] Homeostasis during Development (A) Immunoblots (representative of 3–5 mice) show the expression of VGLUT1–3, synaptophysin (SyPh) and SV2A, GAPDH, and NaPi transporters PiT1 and 2 in murine synaptosomal preparations from developing (E18, postnatal day 3 [P3], and P5) or adult (P90) wild-type mice (left) or from E18 genotypes of the VGLUT2 mouse strains (right). (B) Pi content of E18 synaptosomes obtained from the VGLUT2 −/− strain was drastically reduced in the absence of VGLUTs. (C) (Left) Normalized data from the experiments displayed in (B) show a genotype-specific decline of Pi uptake. (Right) Normalized data from (B) present the RB-mediated decrease in cytosolic Pi for each genotype as % of the control condition. In the absence of Pi, VGLUT activity restores Pi loss, a process inhibited by RB. No restoration of lost Pi takes place in the absence of any VGLUT, and RB has no effect. Values represent pmol [Pi]/μg of protein for each genotype (B) or % of the control condition (no Pi offered; C). Data are the mean ± SEM from at least 6 mice per genotype. Significance toward: (1) no Pi added; (5) VGLUT2 KO control; (6) VGLUT2 WT; and (7) VGLUT2 Het.

Article Snippet: Rabbit PiT1 (SLC20A1) , Gene Tex , GTX64727.

Techniques: Western Blot, Expressing, Control, Activity Assay

Journal: Cell Reports

Article Title: Vesicular Glutamate Transporters (SLCA17 A6, 7, 8) Control Synaptic Phosphate Levels

doi: 10.1016/j.celrep.2020.108623

Figure Lengend Snippet:

Article Snippet: Rabbit PiT1 (SLC20A1) , Gene Tex , GTX64727.

Techniques: Plasmid Preparation, Virus, Recombinant, Clone Assay, Cotransfection

Journal: Cell Reports

Article Title: Vesicular Glutamate Transporters (SLCA17 A6, 7, 8) Control Synaptic Phosphate Levels

doi: 10.1016/j.celrep.2020.108623

Figure Lengend Snippet:

Article Snippet: Rabbit PiT1 (SLC20A1) , Gene Tex , GTX64727.

Techniques: Western Blot