Journal: Neurotherapeutics

Article Title: Continuous intravenous infusion of human mesenchymal stromal cell-derived small extracellular vesicles in spinal cord injured rat modulates extracellular matrix and has greater therapeutic efficacy than multiple single injections

doi: 10.1016/j.neurot.2026.e00892

Figure Lengend Snippet: Characterization of hMSC-sEVs and experimental timelines. (a) Transmission electron micrograph of hMSC-sEVs showing typical morphology. Scale bar = 100 nm. (b) Size distribution of hMSC-sEVs, with most vesicles between 80 and 200 nm. (c) Western blot analysis of three independent hMSC-sEV preparations and their corresponding parental hMSCs. Exosomal markers Alix, CD63, and CD9 were enriched in hMSC-sEVs relative to hMSCs. The endoplasmic reticulum marker Calnexin was strongly detected in hMSC lysates but showed only minimal expression in hMSC-sEV preparations, confirming negligible contamination with intracellular organelle-derived material. (d, e) Treatment timelines for 3-day (d) and 6-day (e) protocols starting on Day 7 post-SCI. Rats received (1) Daily PBS injections, (2) PBS administered via osmotic pump, (3) daily injections of human MSC-derived sEVs(hMSC-sEVs), and (4) hMSC-sEVs administered via osmotic pump. Treatments began on Day 7 post-SCI and continued for 3 (d) or 6 (e) days. Functional recovery (BBB score) and body weight were monitored for 10 weeks; tissues were collected on Day 70 for IHC and western blot. Abbreviations: hMSC-sEVs, human mesenchymal stem cell-derived small extracellular vesicles; BBB, Basso, Beattie, and Bresnahan locomotor score; IHC, immunohistochemistry; WB, western blot; PBS, phosphate-buffered saline; SCI, spinal cord injury.

Article Snippet: To prepare cells for extracellular vesicle (EV) isolation, hMSCs were washed three times with phosphate-buffered saline (PBS, Thermo Fisher) to remove residual FCS-derived vesicles and cultured in serum-free DMEM for 48 h. The conditioned medium was then harvested and subjected to differential ultracentrifugation for small EV (sEV) isolation, following MISEV2023 recommendations [ ] and the protocol by Théry et al., 2006 [ ].

Techniques: Transmission Assay, Western Blot, Marker, Expressing, Derivative Assay, Functional Assay, Immunohistochemistry, Saline

Journal: Neurotherapeutics

Article Title: Continuous intravenous infusion of human mesenchymal stromal cell-derived small extracellular vesicles in spinal cord injured rat modulates extracellular matrix and has greater therapeutic efficacy than multiple single injections

doi: 10.1016/j.neurot.2026.e00892

Figure Lengend Snippet: Effects of daily injections or continuous infusion of hMSC-sEVs on locomotor recovery after SCI. (a–b) Time course of BBB locomotor scores for SCI rats treated with (hMSC-sEVs) or PBS over 3 days (a) or 6 days (b). Animals received daily injections (blue symbols) or continuous pump infusion (red symbols) of hMSC-sEVs beginning at 7 days post-SCI. Grey diamonds represent PBS-treated groups (pooled data from daily injection and pump infusion conditions). Dashed lines at 8 and 10 were added to facilitate comparisons between final BBB scores for daily injection and pump infusion conditions in 3-day and 6-day treatment protocols. n = 15 for all hMSC-sEVs treatment groups, n = 7 or 8 for individual PBS treatment groups, and n = 15 for combined PBS groups (daily injection and pump infusion) with either 3-day or 6-day treatment. (c–e) Bar graphs Comparing BBB scores at: 14 days (c), 21 days (d), and 70 days (e) post-SCI. At 14 and 21 days post-SCI. Statistical significance: #p < 0.05, ##p < 0.01, ###p < 0.001 for comparisons between daily and continuous treatment groups (Daily vs. Pump); ◇◇p < 0.01, ◇◇◇p < 0.001 for comparisons between daily MSC-sEVs and PBS-treated groups; ◆◆◆p < 0.001 for comparisons between continuous MSC-sEVs and PBS-treated groups; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 for comparisons between other treatment groups. (f) Images showing typical body postures of locomoting rats at 70 post SCI after treatment with daily PBS injections, continuous PBS infusion, daily hMSC-sEVs injections, or continuous hMSC-sEVs infusion for 6 days. Abbreviations: SCI: Spinal Cord Injury, PBS: Phosphate-Buffered Saline, MSC: Mesenchymal Stem Cell, sEVs: Small Extracellular Vesicles, hMSC-sEVs: Human Mesenchymal Stem Cell-Derived Small Extracellular Vesicles, BBB score: Basso, Beattie, and Bresnahan locomotor rating scale.

Article Snippet: To prepare cells for extracellular vesicle (EV) isolation, hMSCs were washed three times with phosphate-buffered saline (PBS, Thermo Fisher) to remove residual FCS-derived vesicles and cultured in serum-free DMEM for 48 h. The conditioned medium was then harvested and subjected to differential ultracentrifugation for small EV (sEV) isolation, following MISEV2023 recommendations [ ] and the protocol by Théry et al., 2006 [ ].

Techniques: Injection, Saline, Derivative Assay

Journal: Neurotherapeutics

Article Title: Continuous intravenous infusion of human mesenchymal stromal cell-derived small extracellular vesicles in spinal cord injured rat modulates extracellular matrix and has greater therapeutic efficacy than multiple single injections

doi: 10.1016/j.neurot.2026.e00892

Figure Lengend Snippet: Recovery of growth trajectory after SCI. (a): Representative images showing the gross appearance rats 70 days post- SCI, following 3-day treatment or 6-day treatment with PBS vehicle alone or hMSC-sEVs in PBS. Scale bar indicates 3 cm. (i–j): Line graphs of body weight in 3-day treatment (b) and 6-day treatment (c). Dashed lines at 300 and 350 g indicate approximate weights of 3-day PBS treated and 3 day hMSC-sEVs treated groups at 70 days post SCI and are included as references to compare 3 day and 6 day treatment conditions. (d, e): Bar graphs comparing the body weight (d) and head-body lengths (e) of PBS or hMSC-sEVs treated rats 70 days post-SCI. n = 15 for all hMSC-sEVs treatment groups and 7 or 8 for individual PBS treatment groups and n = 15 for combined daily injection and pump PBS groups with 3-day or 6-day treatment. Statistical significance: ◇p < 0.05, ◇◇p < 0.01, ◇◇◇p < 0.001 for comparisons between daily MSC-sEVs and PBS-treated groups; ◆p < 0.05, ◆◆p < 0.01, ◆◆◆p < 0.001 for comparisons between continuous MSC-sEVs and PBS-treated groups. Abbreviations: SCI: Spinal Cord Injury, PBS: Phosphate-Buffered Saline, hMSC-sEVs: Human Mesenchymal Stem Cell-Derived Small Extracellular Vesicles, n: Sample Number.

Article Snippet: To prepare cells for extracellular vesicle (EV) isolation, hMSCs were washed three times with phosphate-buffered saline (PBS, Thermo Fisher) to remove residual FCS-derived vesicles and cultured in serum-free DMEM for 48 h. The conditioned medium was then harvested and subjected to differential ultracentrifugation for small EV (sEV) isolation, following MISEV2023 recommendations [ ] and the protocol by Théry et al., 2006 [ ].

Techniques: Injection, Saline, Derivative Assay

Journal: Neurotherapeutics

Article Title: Continuous intravenous infusion of human mesenchymal stromal cell-derived small extracellular vesicles in spinal cord injured rat modulates extracellular matrix and has greater therapeutic efficacy than multiple single injections

doi: 10.1016/j.neurot.2026.e00892

Figure Lengend Snippet: Expression of Collagen and Fibronectin at the lesion site. (a) Fluorescence micrographs showing lesion sites in representative animals treated with PBS or hMSC-sEVs at 14 days post-SCI, stained with CD206 (red) and fibronectin (green). Enlarged views of boxed areas from a and b, highlighting the co-localization of CD206 and fibronectin. (b) Fluorescence micrographs showing lesion sites in representative animals treated with PBS or hMSC-sEVs at 70 days post-SCI, stained with CD206 (red), fibronectin (green), and collagen 1 (magenta). (c) Western blot analysis of fibronectin, collagen 1, and collagen 5 levels in spinal cord segments centered around the lesion site at 70 days post-SCI, comparing PBS and hMSC-sEVs treated animals. (d–f) Quantitative analysis of western blot data for fibronectin (d), collagen 1 (e), and collagen 5 (f) normalized to control levels, showing significant reductions in extracellular matrix protein deposition in hMSC-sEV-treated groups compared to PBS-treated groups. Scale bar in lower right a = 20 μm and applies to all other high magnification images. Scale bars in upper right a and lower right b = 100 μm and apply to all low magnification images. Boxed areas marked with an asterisk (∗) in the top row indicate the regions shown at higher magnification in the bottom rowStatistical significance: ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. ∗∗∗∗p < 0.0001. PBS: Phosphate-Buffered Saline, Abbreviations: hMSC-sEVs: Human Mesenchymal Stem Cell-Derived Small Extracellular Vesicles, SCI: Spinal Cord Injury, CD206: Cluster of Differentiation 206.

Article Snippet: To prepare cells for extracellular vesicle (EV) isolation, hMSCs were washed three times with phosphate-buffered saline (PBS, Thermo Fisher) to remove residual FCS-derived vesicles and cultured in serum-free DMEM for 48 h. The conditioned medium was then harvested and subjected to differential ultracentrifugation for small EV (sEV) isolation, following MISEV2023 recommendations [ ] and the protocol by Théry et al., 2006 [ ].

Techniques: Expressing, Fluorescence, Staining, Western Blot, Control, Saline, Derivative Assay

Journal: Nature Communications

Article Title: PRL3-zumab as an immunotherapy to inhibit tumors expressing PRL3 oncoprotein

doi: 10.1038/s41467-019-10127-x

Figure Lengend Snippet: PRL3-zumab inhibits PRL3 + liver tumors in vivo but not cancer cells in vitro. a Representative western blot (WB) of PRL3 protein expression in human (lanes 1–6) and murine (lanes 7 and 8) liver cancer cells. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as a loading control. Asterisks indicate cell lines that rapidly generate orthotopic liver tumors within 5 weeks. b Outline of orthotopic “seed and soil” liver tumor model for treatments. c – e Mean volumes at the end of the experiment in treated (filled squares) and untreated (filled triangles) groups of mice bearing PRL3 + MHCC-LM3 tumors ( n = 10 mice per group; c ), PRL3 − Hep53.4 tumors ( n = 3 mice per group; d ), and Hep53.4-PRL3 tumors ( n = 6 mice per group; e ). The mean value was calculated by the Student’s t test (mean ± s.e.m.). P values between treatment pairs as indicated. Lower panels, representative liver tumors at the end of experiment. Scale bar, 10 mm. f – h The viabilities of MHCC-LM3 cells ( f ), Hep53.4 cells ( g ), and Hep53.4-PRL3 cells ( h ) cultured for 48 h with PBS control (filled squares), 5 µg mL −1 PRL3-zumab (filled upright triangles), 50 µg mL −1 PRL3-zumab (filled inverted triangles), 2 µg mL −1 cisplatin (filled diamonds), or 10 µg mL −1 cisplatin (filled circles) were evaluated by an MTS (3-(4,5-dimethylthiazol-2-yl)−5-(3-carboxymethoxyphenyl)−2-(4-sulfophenyl)-2 H -tetrazolium) assay. The mean value was calculated by the Student’s t test (mean ± s.e.m., n = 3 biologically independent samples each). * P < 0.05, ** P < 0.01, NS, not significant, as compared between treatment and control group for each cell line. Source data are provided as a Source Data file

Article Snippet: The Hep53.4-PRL3 cell line, which stably expresses EGFP-tagged PRL3 fusion protein, was generated by using jetPRIME reagent (Polyplus-transfection) to transfect Hep53.4 cells with the pEGFP-C1-PRL-3 plasmid .

Techniques: In Vivo, In Vitro, Western Blot, Expressing, Cell Culture

Journal: Nature Communications

Article Title: PRL3-zumab as an immunotherapy to inhibit tumors expressing PRL3 oncoprotein

doi: 10.1038/s41467-019-10127-x

Figure Lengend Snippet: Intracellular PRL3 is externalized for PRL3-zumab binding. a Methodology for cell surface analysis of MHCC-LM3 cultured cells (CC) and tumor cells. b – d “Surface” detection of nonspecific control antigens ( b ), EGFR ( c ), or PRL3 ( d ) were detected by fluorescence-activated cell sorting (FACS) analysis using polyclonal human hIgG, anti-EGFR antibody (cetuximab), or anti-PRL3 antibody (PRL3-zumab), respectively. Representative FACS profiles from four biological replicates are shown. e Mean percentage ± s.d. of surface positive (surface+) live cells for each antigen were calculated by dividing the surface antigen-positive live cells (upper left quadrant) by total live cells (sum of both upper and lower left quadrants) in b – d . f Background-corrected values from e were normalized to CC surface expression levels for EGFR (filled circles) and PRL3 (filled squares). The mean fold change was calculated by the Student’s t test (mean ± s.d., n = 4 biologically independent samples). P values as indicated for each antigen. g Background-corrected values of MHCC-LM3 cells cultured under “Normal” vs. “Serum-starved” conditions for 72 h were normalized to “Normal” surface+ cell percentages for each antigen. The mean fold-change was calculated by the Student’s t test (mean ± s.d.) for EGFR (filled circles; n = 3 independent samples) and PRL3 (filled squares; n = 4 independent samples). P values as indicated for each antigen. Source data are provided as a Source Data file

Article Snippet: The Hep53.4-PRL3 cell line, which stably expresses EGFP-tagged PRL3 fusion protein, was generated by using jetPRIME reagent (Polyplus-transfection) to transfect Hep53.4 cells with the pEGFP-C1-PRL-3 plasmid .

Techniques: Binding Assay, Cell Culture, Fluorescence, FACS, Expressing

Journal: Nature Communications

Article Title: PRL3-zumab as an immunotherapy to inhibit tumors expressing PRL3 oncoprotein

doi: 10.1038/s41467-019-10127-x

Figure Lengend Snippet: PRL3 is expressed on the surface of exosomes from PRL3 + cancer cells. a Representative PRL3 western blot (WB) in purified exosomes from MHCC-LM3 and Hep53.4 liver cancer cells cultured for 24 h in serum-free media. Paxillin served as a negative marker for exosomes, whereas TSG101 is a positive exosomal marker. b Immunofluorescence analysis of MHCC-LM3 cells transiently transfected with green fluorescence protein (GFP) or GFP-PRL3 to reveal plasma membrane enrichment of PRL3. Scale bar, 20 µm. c Representative PRL3 WB in purified exosomes from cells in b after culturing for 24 h in serum-free media. Calnexin served as a negative marker for exosomes. d Purified exosomes from Hep53.4 and Hep53.4-PRL3 were analyzed as in c . e Purified exosomes from cells in c , d were assayed for sensitivity of exosomal PRL3 to proteinase K (Prot. K) digestion in the absence or presence of 1% Triton X-100 detergent. TG101 and Alix are intravesicular exosomal proteins. f Proposed model of the localization of PRL3, TSG101, and Alix in exosomes, based on Prot. K susceptibility observed in e . Source data are provided as a Source Data file

Article Snippet: The Hep53.4-PRL3 cell line, which stably expresses EGFP-tagged PRL3 fusion protein, was generated by using jetPRIME reagent (Polyplus-transfection) to transfect Hep53.4 cells with the pEGFP-C1-PRL-3 plasmid .

Techniques: Western Blot, Purification, Cell Culture, Marker, Immunofluorescence, Transfection, Fluorescence, Membrane

Journal: Nature Communications

Article Title: PRL3-zumab as an immunotherapy to inhibit tumors expressing PRL3 oncoprotein

doi: 10.1038/s41467-019-10127-x

Figure Lengend Snippet: PRL3-zumab eliminates tumors in an Fc- and FcR-dependent manner. a Cartoon depicting domain architecture of PRL3-zumab (intact Fc) vs. PRL3-minibody (truncated Fc lacking CH1 and CH2 domains) and their ability to engage Fc receptors (FcR) on host immune cells. The anti-CD16/32 FcR blocker antibody prevents IgG from binding murine FcγII/III receptors. b Both Fc region of PRL3-zumab and FcR binding are required for anti-tumor effects of PRL3-zumab. Upper panel, representative images of livers from each treatment group at day 35 (5-week endpoint). Tumor areas are framed with black lines. Scale bar, 10 mm. Lower panel, tumor volumes in each group. The mean tumor volumes were calculated using one-way analysis of variance (ANOVA) (mean ± s.e.m.) for independent groups of untreated ( n = 12), human IgG-treated ( n = 3), PRL3-zumab-treated ( n = 6), PRL3-minibody-treated ( n = 5), PRL3-zumab + FcR blocked-treated ( n = 7), and FcR blocker-treated ( n = 4) mice. Source data are provided as a Source Data file

Article Snippet: The Hep53.4-PRL3 cell line, which stably expresses EGFP-tagged PRL3 fusion protein, was generated by using jetPRIME reagent (Polyplus-transfection) to transfect Hep53.4 cells with the pEGFP-C1-PRL-3 plasmid .

Techniques: Binding Assay

Journal: Nature Communications

Article Title: PRL3-zumab as an immunotherapy to inhibit tumors expressing PRL3 oncoprotein

doi: 10.1038/s41467-019-10127-x

Figure Lengend Snippet: PRL3-zumab recruits immune cells to tumor sites in an FcR-dependent manner. a – c Orthotopic MHCC-LM3 liver tumor tissue cryo-sections from various groups of mice: I) untreated (filled black squares), II) PRL3-zumab treatment (filled red triangles), III) FcR blocker treatment (filled black triangles), and IV) PRL3-zumab plus Fc blocker combination treatment (filled black diamonds) were analyzed by immunofluorescence with antibodies against B220/CD45R (B cells; a ), CD335 (NK cells; b ), or F4/80 (macrophages; c ) and scored for relative tumor infiltration. The mean relative infiltration was calculated using one-way analysis of variance (ANOVA) (mean ± s.e.m., n = 4 independent samples). Lower panels, representative immunofluorescence results for each antibody set. Scale bar, 200 µm. d Proposed mechanism of action of PRL3-zumab. Externalized PRL3 antigens are recognized by PRL3-zumab, which then recruits NK cells, B cells, and Ly-6C + F4/80 + macrophages for tumor killing and leakage of additional PRL3 antigens (“kill-and-leak” cycle) triggering an anti-tumor ADCC/ADCP cascade. Source data are provided as a Source Data file

Article Snippet: The Hep53.4-PRL3 cell line, which stably expresses EGFP-tagged PRL3 fusion protein, was generated by using jetPRIME reagent (Polyplus-transfection) to transfect Hep53.4 cells with the pEGFP-C1-PRL-3 plasmid .

Techniques: Immunofluorescence

Journal: Nature Communications

Article Title: PRL3-zumab as an immunotherapy to inhibit tumors expressing PRL3 oncoprotein

doi: 10.1038/s41467-019-10127-x

Figure Lengend Snippet: PRL3 is frequently overexpressed in multiple human cancers. a – k Representative full western blot (WB) of PRL3 protein levels in tumor (“T”) tissues and, where available, patient-matched normal (“n”) tissues from a liver, b lung, c colon, d breast, e stomach, f thyroid, g pancreas, h kidney, i acute myeloid leukemia (bone marrow aspirates), j bladder, and k prostate tissues. Relative molecular masses (in kDa) are indicated on the right of each immunoblot. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as a loading control. Source data are provided as a Source Data file

Article Snippet: The Hep53.4-PRL3 cell line, which stably expresses EGFP-tagged PRL3 fusion protein, was generated by using jetPRIME reagent (Polyplus-transfection) to transfect Hep53.4 cells with the pEGFP-C1-PRL-3 plasmid .

Techniques: Western Blot

Journal: Nature Communications

Article Title: PRL3-zumab as an immunotherapy to inhibit tumors expressing PRL3 oncoprotein

doi: 10.1038/s41467-019-10127-x

Figure Lengend Snippet: Summary of PRL3 expression across different tumor types

Article Snippet: The Hep53.4-PRL3 cell line, which stably expresses EGFP-tagged PRL3 fusion protein, was generated by using jetPRIME reagent (Polyplus-transfection) to transfect Hep53.4 cells with the pEGFP-C1-PRL-3 plasmid .

Techniques: Expressing

Journal:

Article Title: Functional inhibition of native volume-sensitive outwardly rectifying anion channels in muscle cells and Xenopus oocytes by anti-ClC-3 antibody

doi: 10.1111/j.1469-7793.2001.0437i.x

Figure Lengend Snippet: A, phase contrast (left panel) and fluorescence micrographs (right panel) of NIH/3T3 cell transfected with gpClC-3-GFP. B, representative IgpClC-3 recorded from GFP-positive NIH/3T3 cells (as shown in A) under isotonic, hypotonic and hypertonic conditions over the range -100 to +120 mV. C, effects of anti-ClC-3 Ab on IgpClC-3 at ±80 mV when pipette solutions contained preabsorbed anti-ClC-3 Ab (control, ○) or anti-ClC-3 Ab alone (▪). D, mean current densities recorded from untransfected NIH/3T3 cells (blue, n = 38), gpClC-3-GFP-transfected cells dialysed with standard intracellular pipette solutions (green, n = 9), preabsorbed anti-ClC-3 Ab (black, n = 4), and anti-ClC-3 Ab alone (red, n = 9).

Article Snippet: Furthermore, in cardiac tissues endogenous ClC-3 protein expression has been confirmed using the same anti-ClC-3 Ab (Alomone Labs) in immunoblot and immunofluorescence experiments ( Britton et al. 2000 ). and illustrate the effects of intracellular dialysis of anti-ClC-3 Ab alone or antigen-preabsorbed anti-ClC-3 Ab on native VSOACs in guinea-pig cardiac and canine pulmonary arterial smooth muscle myocytes, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 2 caption a7 caption a8 Anti-ClC-3 Ab dialysis abolishes native VSOAC currents in guinea-pig cardiac myocytes A , native VSOAC currents at ±80 mV in myocytes exposed to isotonic and hypotonic solutions.

Techniques: Fluorescence, Transfection, Transferring

Journal:

Article Title: Functional inhibition of native volume-sensitive outwardly rectifying anion channels in muscle cells and Xenopus oocytes by anti-ClC-3 antibody

doi: 10.1111/j.1469-7793.2001.0437i.x

Figure Lengend Snippet: A, time course of VSOAC currents from two PASMCs dialysed with either anti-ClC-3 Ab (5 μg ml−1) preabsorbed with antigen (50 μg ml−1; ○), or anti-ClC-3 Ab alone (5 μg ml−1; •). Inset: Western blot analysis of native ClC-3 expression in isolated canine PASMCs. B, mean current densities in cells dialysed with either standard intracellular solutions (n = 11), preabsorbed anti-ClC-3 Ab (n = 4) or anti-ClC-3 Ab alone (n = 5).

Article Snippet: Furthermore, in cardiac tissues endogenous ClC-3 protein expression has been confirmed using the same anti-ClC-3 Ab (Alomone Labs) in immunoblot and immunofluorescence experiments ( Britton et al. 2000 ). and illustrate the effects of intracellular dialysis of anti-ClC-3 Ab alone or antigen-preabsorbed anti-ClC-3 Ab on native VSOACs in guinea-pig cardiac and canine pulmonary arterial smooth muscle myocytes, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 2 caption a7 caption a8 Anti-ClC-3 Ab dialysis abolishes native VSOAC currents in guinea-pig cardiac myocytes A , native VSOAC currents at ±80 mV in myocytes exposed to isotonic and hypotonic solutions.

Techniques: Western Blot, Expressing, Isolation

Journal:

Article Title: Functional inhibition of native volume-sensitive outwardly rectifying anion channels in muscle cells and Xenopus oocytes by anti-ClC-3 antibody

doi: 10.1111/j.1469-7793.2001.0437i.x

Figure Lengend Snippet: A, native VSOAC currents at ±80 mV in myocytes exposed to isotonic and hypotonic solutions. Aa, pipette solutions contained preabsorbed anti-ClC-3 Ab (control, ○) or anti-ClC-3 Ab alone (▪). Ab, mean current densities for preabsorbed anti-ClC-3 Ab (□, n = 3) or anti-ClC-3 Ab alone (▪, n = 5) at times indicated. B, same as A, except cells were preswelled by pre-exposure to hypotonic bath solutions prior to membrane rupture and dialysis; preabsorbed anti-ClC-3 Ab (□, n = 4), and anti-ClC-3 Ab alone (▪, n = 5).

Article Snippet: Furthermore, in cardiac tissues endogenous ClC-3 protein expression has been confirmed using the same anti-ClC-3 Ab (Alomone Labs) in immunoblot and immunofluorescence experiments ( Britton et al. 2000 ). and illustrate the effects of intracellular dialysis of anti-ClC-3 Ab alone or antigen-preabsorbed anti-ClC-3 Ab on native VSOACs in guinea-pig cardiac and canine pulmonary arterial smooth muscle myocytes, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 2 caption a7 caption a8 Anti-ClC-3 Ab dialysis abolishes native VSOAC currents in guinea-pig cardiac myocytes A , native VSOAC currents at ±80 mV in myocytes exposed to isotonic and hypotonic solutions.

Techniques: Transferring

Journal:

Article Title: Functional inhibition of native volume-sensitive outwardly rectifying anion channels in muscle cells and Xenopus oocytes by anti-ClC-3 antibody

doi: 10.1111/j.1469-7793.2001.0437i.x

Figure Lengend Snippet: A, Western blot analysis of native ClC-3 expression in oocytes. B, 4-phorbol 12,13-dibutyrate (PDBu) inhibition of native VSOAC currents in oocytes (-100 to +120 mV). C, time course of VSOAC activation in response to hypotonic bath solutions from a non-injected control oocyte (○) and an oocyte injected with anti-ClC-3 Ab (15 μg ml-1; •) at time shown. Cells were held at -30 mV for 30 ms, hyperpolarized to -100 mV for 210 ms and then depolarized to +100 mV for 210 ms, repetitively at 2 Hz. D, mean current densities from control oocytes (n = 10), oocytes injected with anti-ClC-3 Ab alone (15 μg ml-1) (n = 4) or injected with preabsorbed anti-ClC-3 Ab (150 μg ml-1) (n = 4). Peak current densities were measured after 5 min exposure to isotonic solutions, and after 85 min exposure to hypotonic solutions.

Article Snippet: Furthermore, in cardiac tissues endogenous ClC-3 protein expression has been confirmed using the same anti-ClC-3 Ab (Alomone Labs) in immunoblot and immunofluorescence experiments ( Britton et al. 2000 ). and illustrate the effects of intracellular dialysis of anti-ClC-3 Ab alone or antigen-preabsorbed anti-ClC-3 Ab on native VSOACs in guinea-pig cardiac and canine pulmonary arterial smooth muscle myocytes, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 2 caption a7 caption a8 Anti-ClC-3 Ab dialysis abolishes native VSOAC currents in guinea-pig cardiac myocytes A , native VSOAC currents at ±80 mV in myocytes exposed to isotonic and hypotonic solutions.

Techniques: Western Blot, Expressing, Inhibition, Activation Assay, Injection