Journal:

Article Title: Effects of CXCR4 Gene Transfer on Cardiac Function After Ischemia-Reperfusion Injury

doi: 10.2353/ajpath.2010.090451

Figure Lengend Snippet: CXCL12 is upregulated in ischemic myocardium of CXCR4-overexpressed rat. A: CXCL12 protein expression was assessed in hearts injected with either CXCR4 or β-gal and/or control (saline-injected; with IR) by immunofluorescence staining. Frozen sections were fixed and stained with anti-CXCL12 (green) and anti–α actinin (red). Primary Abs visualized with FITC or Texas Red conjugate. Nuclei were stained with DAPI. Images are taken with confocal microscopy. B: Protein lysates were prepared from rats’ hearts one week after CXCR4 gene transfer followed by 30 minutes LAD ligation and 24 hours reperfusion. The ischemic and remote tissues were lysed and analyzed by Western blot. Representative gel of three independent experiments is shown. C: CXCL12 mRNA levels were determined by quantitative real-time PCR (QRT-PCR) using a QuantiTect SYBR Green RT-PCR Kit and using specific primers for CXCL12 and 18S. Primers were designed to generate short amplification products. Densitometric analysis of data from three different experiments is shown. *P < 0.05.

Article Snippet: The following antibodies were used: (1) rabbit anti-mouse/rat CXCL12 (eBiosciences; San Diego, CA); (2) goat anti-rat CXCR4 antibody (Torrey Pines Biolabs; East Orange, NJ); (3) goat anti-mouse/rat TNF- α, (R&D), (4) mouse anti-rat leukocyte antibody (BD Pharmingen; Franklin Lakes, NJ); and (5) mouse anti- rat macrophage marker (VP-M640, Vector Laboratories; Burlingame, CA) Sections from the hearts were fixed and blocked with the normal serum and incubated with primary antibody for 2 hours at room temperature.

Techniques: Expressing, Injection, Immunofluorescence, Staining, Confocal Microscopy, Ligation, Western Blot, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction, Amplification

Journal: Journal of pharmacological sciences

Article Title: HMGB1 and its membrane receptors as therapeutic targets in an intravesical substance P-induced bladder pain syndrome mouse model.

doi: 10.1016/j.jphs.2020.03.002

Figure Lengend Snippet: Fig. 3. Plasma HMGB1 levels and expression of HMGB1, RAGE, TLR4, CXCR4 or CXCL12 in the bladder tissue after intravesical substance P in mice. Blood collection and bladder excision were performed 24 h after intravesical substance P (SP). V, vehicle. Plasma HMGB1 levels were determined by ELISA, and protein levels were quantified by Western blotting. V, vehicle. Data show the mean with S.E.M. for 5e6 (A), 4 (B), 7 (C, D) or 6 (E, F) mice. *P < 0.05 vs V.

Article Snippet: *P < 0.05, **P < 0.01, ***P < 0.001 vs V þ V; yP < 0.05, yyP < 0.01 vs IgG Biotechnology, Santa Cruz, CA, USA), anti-RAGE rabbit polyclonal antibody (1:2000 dilution) (Abcam, Cambridge, UK), anti-TLR4 rabbit polyclonal antibody (1:200 dilution) (Santa Cruz Biotechnology), the anti-CXCR4 rabbit polyclonal antibody (1:5000 dilution) (NOVUSBIOLOGICALS, Littleton, CO, USA), and anti-SDF1 (CXCL12) rabbit polyclonal antibody (1:3000 dilution) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Clinical Proteomics, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: PLoS ONE

Article Title: High-mobility group box 1 fragment suppresses adverse post-infarction remodeling by recruiting PDGFRα-positive bone marrow cells

doi: 10.1371/journal.pone.0230392

Figure Lengend Snippet: The second examination was performed using the same MI rats prior to HMGB1 treatment to assess the expression of SDF1, a representative homing factor of MSCs. A: Study protocol of second examination. B: Histological analysis revealed SDF1 expression along the peri-infarction area in MI model and normal rats (40×, scale bar = 200 μm). C: RT-PCR analysis indicated that SDF1 expression increased significantly in MI rats (n = 6) compared with normal rats (n = 10). P -values were calculated using the Welch’s t-test. P < 0.05*, P < 0.01**.

Article Snippet: In the second examination, the frozen sections in MI and normal rats were stained with rabbit anti-SDF1 polyclonal antibody (1:50; Abcam, Cambridge, UK), and were evaluated using the confocal laser microscopy.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: High-mobility group box 1 fragment suppresses adverse post-infarction remodeling by recruiting PDGFRα-positive bone marrow cells

doi: 10.1371/journal.pone.0230392

Figure Lengend Snippet: Intravital imaging analysis was performed using the GFP-BMT rat MI model to visualize HMGB1-induced GFP + -cells mobilization to the damaged heart tissue in real time. A: Details of the study protocol using intravital imaging. B: Histological findings in GFP-BMT rat MI model 12 h after HMGB1 treatment. 1, 2) More GFP + /PDGFRα + cells were visualized in the HMGB1 group compared with the control (100×, 600×; scale bar = 100, 200 μm, respectively). C: In the HMGB1 group, GFP + -cells were recruited along the peri-infarction area with SDF1 over-expression (100×, scale bar = 200 μm). Conversely, the recruitment of GFP + -cells was not enhanced at the remote area, where significant expression of SDF1 was not observed. P -values were calculated using the Welch’s t-test. P < 0.01**.

Article Snippet: In the second examination, the frozen sections in MI and normal rats were stained with rabbit anti-SDF1 polyclonal antibody (1:50; Abcam, Cambridge, UK), and were evaluated using the confocal laser microscopy.

Techniques: Imaging, Over Expression, Expressing

Journal: PLoS ONE

Article Title: High-mobility group box 1 fragment suppresses adverse post-infarction remodeling by recruiting PDGFRα-positive bone marrow cells

doi: 10.1371/journal.pone.0230392

Figure Lengend Snippet: Systemic administration of HMGB1 fragment can mobilize BM mesenchymal cells, including BM-MSCs, to blood circulation. Consequently, these BM-derived mesenchymal cells accumulate in the damaged myocardium through the SDF1/CXCR4 signaling complex, leading to functional recovery by paracrine activity of the BM-MSCs or to differentiation of some vascular constituent cells.

Article Snippet: In the second examination, the frozen sections in MI and normal rats were stained with rabbit anti-SDF1 polyclonal antibody (1:50; Abcam, Cambridge, UK), and were evaluated using the confocal laser microscopy.

Techniques: Derivative Assay, Functional Assay, Activity Assay

Journal: eLife

Article Title: Distinct transcriptomic profile of satellite cells contributes to preservation of neuromuscular junctions in extraocular muscles of ALS mice

doi: 10.7554/elife.92644

Figure Lengend Snippet: Figure 8. Coculture of AAV transduced SC-derived myotubes with rat spinal motor neurons in compartmentalized microfluidic chambers. (A) Schematic representation of XonaChip spatial configuration and coculture experiment timeline. NBPM: Neurobasal Plus Medium; RSMNs: rat spinal motor neurons. (B) Representative composite images of RSMNs cocultured with: 1. WT HL SC-derived myotubes transduced with AAV-CMV-eGFP; 2. G93A HL SC-derived myotubes transduced with AAV- CMV-eGFP; 3. G93A HL SC-derived myotubes transduced with AAV-CMV-Cxcl12-IRES-eGFP; 4. G93A EOM

Article Snippet: Peptide, recombinant protein Collagenase II Worthington Cat # LS004176 Final concentration 0.26% Peptide, recombinant protein Dispase II Sigma Aldrich Cat # D46931G Final concentration 0.24% Peptide, recombinant protein Hyaluronidase Worthington Cat # LS002592 Final concentration 0.16% Peptide, recombinant protein DNase I Worthington Cat # LS002139 Final concentration 0.04% Antibody Anti- mouse Vcam1- PE (Rat monoclonal IgG2a) BioLegend Clone 429 (MVCAM.A) Cat # 105713 600 ng/106 cells for FACS isolation Antibody Anti- mouse CD31- APC (Rat monoclonal IgG2a) BioLegend Clone 390 Cat #102409 400 ng/106 cells for FACS isolation Antibody Anti- mouse CD45- APC (Rat monoclonal IgG2b) BioLegend Clone 30- F11 Cat # 103111 400 ng/106 cells for FACS isolation Antibody Anti- mouse Sca- 1- APC (Rat monoclonal IgG2a) BioLegend Clone D7 Cat # 108111 200 ng/106 cells for FACS isolation Antibody Anti- chicken Pax7 (Mouse monoclonal IgG1) Santa Cruz Biotechnology Cat # sc- 81648 IF (1:100) Antibody Anti- mouse MyoD (Mouse monoclonal IgG2b) Santa Cruz Biotechnology Cat # sc- 377460 IF (1:100) Antibody Anti- mouse Ki- 67 (Rabbit monoclonal IgG) Cell Signaling Technology Cat # 12202 S IF (1:300) Antibody Anti- chicken MHC (Mouse monoclonal IgG2b) Developmental Studies Hybridoma Bank Cat # MF20 IF (1:200) Antibody Anti- rat NF- M (Mouse monoclonal IgG1) Developmental Studies Hybridoma Bank Cat # 2H3 IF (1:300) Antibody Anti- GFP (Mouse monoclonal IgG2a) Novus Clone 4B10B2 Cat # NBP222111AF488 IF (1:250) Antibody Anti- rat Synaptophysin (Rabbit polyclonal IgG) Thermo Fisher Scientific PA11043 IF (1:400) Antibody Anti- human Cxcl12 (Mouse monoclonal IgG1) R&D systems Clone 79018 Cat # MAB350 IF (1:200) Other AF 488 conjugated α-Bungarotoxin Thermo Fisher Scientific Cat # B13422 IF (1:1000) Continued Continued on next page Li et al. eLife 2023;12:RP92644.

Techniques: Derivative Assay, Transduction

Journal: eLife

Article Title: Distinct transcriptomic profile of satellite cells contributes to preservation of neuromuscular junctions in extraocular muscles of ALS mice

doi: 10.7554/elife.92644

Figure Lengend Snippet: Figure 9. Coculture of AAV transduced SC-derived myotubes with rat spinal motor neurons within the same compartment. (A) Representative composite images of RSMNs cocultured with: 1. WT HL SC-derived myotubes transduced with AAV-CMV-eGFP; 2. G93A HL SC-derived myotubes transduced with AAV-CMV-eGFP; 3. G93A HL SC-derived myotubes transduced with AAV-CMV- Cxcl12-IRES-eGFP; 4. G93A EOM SC-derived myotubes transduced with AAV-CMV-eGFP. The timeline was the same as that in Figure 8 but RSMNs were seeded on top of myotubes. Boxed regions are

Article Snippet: Peptide, recombinant protein Collagenase II Worthington Cat # LS004176 Final concentration 0.26% Peptide, recombinant protein Dispase II Sigma Aldrich Cat # D46931G Final concentration 0.24% Peptide, recombinant protein Hyaluronidase Worthington Cat # LS002592 Final concentration 0.16% Peptide, recombinant protein DNase I Worthington Cat # LS002139 Final concentration 0.04% Antibody Anti- mouse Vcam1- PE (Rat monoclonal IgG2a) BioLegend Clone 429 (MVCAM.A) Cat # 105713 600 ng/106 cells for FACS isolation Antibody Anti- mouse CD31- APC (Rat monoclonal IgG2a) BioLegend Clone 390 Cat #102409 400 ng/106 cells for FACS isolation Antibody Anti- mouse CD45- APC (Rat monoclonal IgG2b) BioLegend Clone 30- F11 Cat # 103111 400 ng/106 cells for FACS isolation Antibody Anti- mouse Sca- 1- APC (Rat monoclonal IgG2a) BioLegend Clone D7 Cat # 108111 200 ng/106 cells for FACS isolation Antibody Anti- chicken Pax7 (Mouse monoclonal IgG1) Santa Cruz Biotechnology Cat # sc- 81648 IF (1:100) Antibody Anti- mouse MyoD (Mouse monoclonal IgG2b) Santa Cruz Biotechnology Cat # sc- 377460 IF (1:100) Antibody Anti- mouse Ki- 67 (Rabbit monoclonal IgG) Cell Signaling Technology Cat # 12202 S IF (1:300) Antibody Anti- chicken MHC (Mouse monoclonal IgG2b) Developmental Studies Hybridoma Bank Cat # MF20 IF (1:200) Antibody Anti- rat NF- M (Mouse monoclonal IgG1) Developmental Studies Hybridoma Bank Cat # 2H3 IF (1:300) Antibody Anti- GFP (Mouse monoclonal IgG2a) Novus Clone 4B10B2 Cat # NBP222111AF488 IF (1:250) Antibody Anti- rat Synaptophysin (Rabbit polyclonal IgG) Thermo Fisher Scientific PA11043 IF (1:400) Antibody Anti- human Cxcl12 (Mouse monoclonal IgG1) R&D systems Clone 79018 Cat # MAB350 IF (1:200) Other AF 488 conjugated α-Bungarotoxin Thermo Fisher Scientific Cat # B13422 IF (1:1000) Continued Continued on next page Li et al. eLife 2023;12:RP92644.

Techniques: Derivative Assay, Transduction