Journal: eLife

Article Title: Near-perfect precise on-target editing of human hematopoietic stem and progenitor cells

doi: 10.7554/eLife.91288

Figure Lengend Snippet:

Article Snippet: Antibody , FITC anti-human, CD49c (Clone REA360) , Miltenyi , 130-105-364 , 1:50.

Techniques: Plasmid Preparation, Expressing, Sterility, Ligation, Selection, Sequencing, Recombinant, Electroporation, CRISPR, Software

Journal: Investigative ophthalmology & visual science

Article Title: Three-Dimensional Neurite Characterization of Small Incision Lenticule Extraction Derived Lenticules.

doi: 10.1167/iovs.19-27566

Figure Lengend Snippet: FIGURE 4. Interaction between lenticule neurites and SCs. Colocaliza- tion of SC marker expression and stromal neurites by confocal immunofluorescence: (A–C) GAP43 (green) and TuJ1 (red). (D–F) p75NTR (green) and TuJ1 (red).

Article Snippet: Antibodies Used in This Study Antibody Source Working Concentration 1 bIII-tubulin Covance MMS435P 0.5 lg/mL 2 GAP43 Novus NB300-143 0.5 lg/mL 3 P75NTR Millipore AB1554 0.5 lg/mL 4 Phalloidin-AlexaFluor 594 conjugate Invitrogen 0.5 lg/mL 5 AlexaFluor 488 goat anti-mouse/rabbit IgG (HþL) Jackson ImmunoRes Lab Immunostaining: 1:700 6 RedX-conjugated goat anti-mouse/rabbit IgG (HþL) Jackson ImmunoRes Lab Immunostaining: 1:700 Downloaded from iovs.arvojournals.org on 10/24/2019 placed as flat-mount.

Techniques: Marker, Expressing

Journal: Journal of Neuroscience

Article Title: Toll-Like Receptor Signaling Is Critical for Wallerian Degeneration and Functional Recovery after Peripheral Nerve Injury

doi: 10.1523/jneurosci.3027-07.2007

Figure Lengend Snippet: Figure 3. Deficiency in TLR signaling compromises macrophage recruitment/activation and delays myelin debris clearance, axonal regeneration, and locomotor recovery after sciatic nerve lesion. A–D, Representative fluorescence photomicrographs takenfromlongitudinalsectionsofthesciaticnervedistaltothelesionshowingCD68-immunopositivemacrophagesinC57BL/6J (A),TLR2-ko(B),C3H/HeOUJ(C),andTLR4d(D)miceat7d.E,QuantificationofthenumberofCD68macrophagesinthesciaticnerve distalstumpofTLR2-ko,TLR4d,MyD88-ko,andwild-typemiceat7dafterlesion(n8pergroup).F–I,Bright-fieldphotomicrographs showingmyelinstainedwithLFBinthesciaticnervedistalstumpofC57BL/6J(F),TLR2-ko(G),C3H/HeOUJ(H),andTLR4d(I)miceat7d. J,QuantificationofLFBstainingofmyelininthedegeneratingsciaticnervedistalstumpofmicedeficientinTLRsignalingat7d(n8per group). K, Quantification of the number of axons that had regenerated up to 4 mm distal to the site of lesion, as visualized by GAP-43 immunofluorescence, at 4 d after lesion. L, M, Recovery of locomotor functions from a sciatic nerve lesion, as determined by the SFI, in TLR2-ko,MyD88-ko,andTLR4dmicecomparedwiththeirrespectivecontrolgroups(WT)overaperiodof49d(n12pergroup).***p 0.001,**p0.01,and*p0.05comparedwiththecontrolgroup.Scalebar,50m.

Article Snippet: Immunohistochemistry was performed to detect the following antigens: (1) mouse CD68 using the monoclonal anti-CD68 antibody (to visualize activated macrophages/monocytes; Serotec, Raleigh, NC), (2) growth-associated protein-43 (GAP-43) using the polyclonal antiGAP-43 antibody (to visualize regenerating axons; Novus Biologicals, Littleton, CO), (3) rat CD68 using the monoclonal ED-1 antibody (to visualize activated macrophages/monocytes; Serotec), (4) ionized calcium-binding adaptor molecule 1 (iba1) using the polyclonal antiiba1 antibody (to visualize macrophages/monocytes; Wako Chemicals USA, Richmond, VA), and (5) mouse CD45 using the monoclonal antiCD45 antibody (to visualize leukocytes; BD PharMingen, Mississauga, Ontario, Canada).

Techniques: Activation Assay, Fluorescence, Immunofluorescence

Journal: Experimental cell research

Article Title: Dl-3-n-butylphthalide promotes neurite outgrowth of primary cortical neurons by Sonic Hedgehog signaling via upregulating Gap43.

doi: 10.1016/j.yexcr.2020.112420

Figure Lengend Snippet: Figure 4. NBP elevates Shh, Ptch, Gli1 and Gap43 expressions at DIV1. (A)

Article Snippet: The primary antibodies used were as follows: mouse anti-β-tubulin III antibody (TuJ1, 1:500, Sigma), mouse anti-Smi312 antibody (1:500, Convance), mouse anti-Map2 antibody (2a + 2b) (1:500, Sigma), mouse anti-F-actin antibody (1:500, Abcam), rabbit anti-Gap43 antibody (1:5000, Novus), mouse anti-Gap43 antibody (1:5000, Novus), rabbit anti-Shh antibody (1:100, Santa Cruz) and rabbit anti-Ptch antibody (1:1000, Abcam).

Techniques:

Journal: Biology Direct

Article Title: Opening of Cx43-formed hemichannels mediates the Ca 2+ signaling associated with endothelial cell migration

doi: 10.1186/s13062-023-00408-3

Figure Lengend Snippet: Cx43-formed hemichannels, but not gap junction channels, are associated with endothelial cell migration. a , Representative images of dye coupling assay (left) and the analysis of the number of coupled cells via gap junction communication (right) attained in the intact monolayer and in the migration front after scratching the monolayer. Dye coupling was assessed by measuring after 2 min the diffusion to neighboring cells (coupled cells) of the ethidium bromide microinjected into a single endothelial cell. The yellow diamond indicates the microinjected cell. b , Representative images of the ethidium uptake observed in primary cultures of mesenteric endothelial cells in control conditions and in the presence of the Cx blocking peptide 37,43 Gap27 (200 µM) or the Cx43 hemichannel inhibitor TAT-Gap19 (Gap19, 300 µM) (left). Ethidium uptake was evaluated 15 min after scratching the monolayer and cells were incubated with the dye for 15 min, as shown in the time course of ethidium uptake observed in the intact monolayer and in the migration front (b, right top). Dot yellow lines depict the edge of the migration front. In addition, the analysis of the ethidium uptake rate achieved in the intact monolayer and in the migration front in control conditions and in the presence of 37,43 Gap27 or Gap19 is also shown (b, right bottom). The rate of ethidium uptake was assessed by calculating the slope of the increase in fluorescence intensity along the time. Changes in ethidium-fluorescence signal are expressed in arbitrary units (a.u.). Numbers inside the bars indicate the n value. Values are means ± SEM. ***, P < 0.001 vs. Monolayer by two-way ANOVA. ††, P < 0.01 and †††, P < 0.001 vs. Migration front in control conditions (Control) by one-way ANOVA plus Bonferroni post hoc test. &, P < 0.001 vs. Migration front by paired Student’s t-test

Article Snippet: Ascorbic acid was obtained from Santa Cruz Biotechnology (TX, USA) and the peptides 37,43 Gap27 and TAT-Gap19 were from Tocris Bioscience (Bristol, UK).

Techniques: Migration, Diffusion-based Assay, Control, Blocking Assay, Incubation, Fluorescence

Journal: Biology Direct

Article Title: Opening of Cx43-formed hemichannels mediates the Ca 2+ signaling associated with endothelial cell migration

doi: 10.1186/s13062-023-00408-3

Figure Lengend Snippet: Endothelial cell migration depends on a Cx43-formed channel-mediated increase in intracellular Ca 2+ concentration ([Ca 2+ ] i ). a , Representative images (left) and fluorescence intensity analysis (right) of the increase in [Ca 2+ ] i observed in endothelial cells of the migration front 15 min after scratching the monolayer in control conditions and in the presence of 50 µM 18-β-Glycyrrhetenic acid (ß-GA), a general Cx-formed channel blocker, or 200 µM 37,43 Gap27, a peptide designed to block channels formed by Cx37 or Cx43. Variations in the levels of [Ca 2+ ] i were assessed with the fluorescent Ca 2+ indicator Fluo-4. b , Representative images of the changes in [Ca 2+ ] i of endothelial cells (left) in which the differences in the subcellular distribution of the Ca 2+ signal attained in a cell of the migration front (Cell 2) and a cell within the monolayer (Cell 1) are highlighted in a 3D analysis (middle and right). c , Fluorescence intensity analysis of the Fluo-4 signal measured along the endothelial cells length (from back to front) in the migration front and the monolayer. d , Analysis of the changes in [Ca 2+ ] i levels attained in the rear and anterior edge of endothelial cells of the migration front in control conditions and after the treatment with ß-GA or 37,43 Gap27. Changes in Fluo-4 signal are expressed as the area under the curve (AUC). Note that cells were treated with the Cx blocking peptide 37,43 Gap27 for only 10 min to inhibit Cx hemichannels, without affecting gap junction channels. Dot red lines depict the edge of the migration front. Numbers inside the bars or in parentheses indicate the n value. Values are means ± SEM. ***, P < 0.001 vs. Control by one-way ANOVA plus Bonferroni post hoc test. †††, P < 0.001 vs. Anterior by paired Student’s t-test

Article Snippet: Ascorbic acid was obtained from Santa Cruz Biotechnology (TX, USA) and the peptides 37,43 Gap27 and TAT-Gap19 were from Tocris Bioscience (Bristol, UK).

Techniques: Migration, Concentration Assay, Fluorescence, Control, Blocking Assay