Journal: Nature Communications

Article Title: An updated suite of viral vectors for in vivo calcium imaging using intracerebral and retro-orbital injections in male mice

doi: 10.1038/s41467-023-36324-3

Figure Lengend Snippet: List of antibodies used for post-mortem histology

Article Snippet: Donkey anti-goat IgG, CF 568 conj. , Biotium , 1:1000 , AB_10854239 , 20106-1.

Techniques:

Journal: Cancer Science

Article Title: Angiotensin II subtype 1a receptor signaling in resident hepatic macrophages induces liver metastasis formation

doi: 10.1111/cas.13306

Figure Lengend Snippet: Angiotensin II (Ang II )/ AT 1a axis enhances transforming growth factor‐β1 ( TGF ‐β1) expression in Kupffer cells. (a) Accumulation of F4/80 + cells, TGF ‐β1 + cells, and F4/80 + / TGF ‐β1 + cells in metastatic areas from WT and AT 1a KO mice on day 14. Double‐staining of liver sections with antibodies against F4/80 (red) and TGF ‐β1 (green) in WT and AT 1a KO mice. Expression of TGF ‐β1 was colocalized with F4/80 + cells. Metastatic area is delineated with the white dashed line. T, tumor. Scale bar = 100 μm. (b–d) Numbers of F4/80 + cells (b), TGF ‐β1 + cells (c), and F4/80 + TGF ‐β1 + cells (d) in metastatic areas from WT and AT 1a KO mice. Data are expressed as the means ± SD of six mice per group. * P < 0.05. (e) Expression of AT 1a and AT 1b in KUP 5 Kupffer cells. Data are expressed as the means ± SD of six mice per group. (f) Expression of TGF ‐β1 on KUP 5 cells under stimulation with Ang II . Expression of TGF ‐β1 was enhanced 6 h after stimulation with Ang II compared with control. There was no significant difference at 12 h. Data are expressed as the means ± SD of six mice per group. * P < 0.05 versus control.

Article Snippet: For the latter, sections were activated using Histo VT One (Nacalai Tesque, Yokohama, Japan) and then incubated overnight at 4°C with one of the following primary antibodies: (a) anti‐mouse F4/80 antibody (1:200, rat monoclonal, sc52664; Santa Cruz Biotechnology, Dallas, TX, USA); (b) anti‐mouse TGF‐β1 antibody (1:200, rabbit polyclonal, ab92486; Abcam, Cambridge, UK); (c) anti‐mouse GFP antibody (1:200, rabbit polyclonal, ab290; Abcam); (d) anti‐mouse Ang II type 1A receptor antibody (1:100, rabbit polyclonal, bs‐2132R; Bioss, Boston, MA, USA); (e) anti‐mouse desmin antibody (1:100, goat polyclonal, ab80503; Abcam); (f) anti‐mouse type I collagen antibody (1:100, rabbit polyclonal, ab21286; Abcam); or (g) anti‐mouse CD31 antibody (1:200, rabbit polyclonal, ab28364; Abcam).

Techniques: Expressing, Double Staining

Journal: American Journal of Human Genetics

Article Title: Identification of a Kir3.4 Mutation in Congenital Long QT Syndrome

doi: 10.1016/j.ajhg.2010.04.017

Figure Lengend Snippet: The Kir3.4-Gly387Arg Mutation Is Associated with Congenital Long QT Syndrome

Article Snippet: Proteins were transferred onto Hybond-P polyvinylidene difluoride transfer membranes (Amersham Biosciences, 0.45 μm), and membranes were blocked in 5% nonfat milk in TBST (0.1% Tween 20 in Tris-buffered saline solution) for 2 hr at room temperature and then incubated with primary antibody against Kir3.1 (1:1,000; Alomone Labs), Kir3.4 (1:1,000; Alomone Labs), calnexin (1:2,000; Nordic Biosite), or Na + /K+-ATPase α1 (1:500; Santa Cruz Biotechnology) overnight.

Techniques: Mutagenesis

Journal: American Journal of Human Genetics

Article Title: Identification of a Kir3.4 Mutation in Congenital Long QT Syndrome

doi: 10.1016/j.ajhg.2010.04.017

Figure Lengend Snippet: Clinical Characteristics of the Pedigree Members with Kir3.4-Gly387Arg or Sudden Death

Article Snippet: Proteins were transferred onto Hybond-P polyvinylidene difluoride transfer membranes (Amersham Biosciences, 0.45 μm), and membranes were blocked in 5% nonfat milk in TBST (0.1% Tween 20 in Tris-buffered saline solution) for 2 hr at room temperature and then incubated with primary antibody against Kir3.1 (1:1,000; Alomone Labs), Kir3.4 (1:1,000; Alomone Labs), calnexin (1:2,000; Nordic Biosite), or Na + /K+-ATPase α1 (1:500; Santa Cruz Biotechnology) overnight.

Techniques:

Journal: American Journal of Human Genetics

Article Title: Identification of a Kir3.4 Mutation in Congenital Long QT Syndrome

doi: 10.1016/j.ajhg.2010.04.017

Figure Lengend Snippet: Electrophysiological Analyses of Kir3.4-Gly387Arg

Article Snippet: Proteins were transferred onto Hybond-P polyvinylidene difluoride transfer membranes (Amersham Biosciences, 0.45 μm), and membranes were blocked in 5% nonfat milk in TBST (0.1% Tween 20 in Tris-buffered saline solution) for 2 hr at room temperature and then incubated with primary antibody against Kir3.1 (1:1,000; Alomone Labs), Kir3.4 (1:1,000; Alomone Labs), calnexin (1:2,000; Nordic Biosite), or Na + /K+-ATPase α1 (1:500; Santa Cruz Biotechnology) overnight.

Techniques:

Journal: American Journal of Human Genetics

Article Title: Identification of a Kir3.4 Mutation in Congenital Long QT Syndrome

doi: 10.1016/j.ajhg.2010.04.017

Figure Lengend Snippet: Cell Surface Expression of Kir3.4-Gly387Arg and Kir3.1

Article Snippet: Proteins were transferred onto Hybond-P polyvinylidene difluoride transfer membranes (Amersham Biosciences, 0.45 μm), and membranes were blocked in 5% nonfat milk in TBST (0.1% Tween 20 in Tris-buffered saline solution) for 2 hr at room temperature and then incubated with primary antibody against Kir3.1 (1:1,000; Alomone Labs), Kir3.4 (1:1,000; Alomone Labs), calnexin (1:2,000; Nordic Biosite), or Na + /K+-ATPase α1 (1:500; Santa Cruz Biotechnology) overnight.

Techniques: Expressing

Journal: American Journal of Human Genetics

Article Title: Identification of a Kir3.4 Mutation in Congenital Long QT Syndrome

doi: 10.1016/j.ajhg.2010.04.017

Figure Lengend Snippet: Human Atrial and Ventricular Expression of Kir3.1 and Kir3.4

Article Snippet: Proteins were transferred onto Hybond-P polyvinylidene difluoride transfer membranes (Amersham Biosciences, 0.45 μm), and membranes were blocked in 5% nonfat milk in TBST (0.1% Tween 20 in Tris-buffered saline solution) for 2 hr at room temperature and then incubated with primary antibody against Kir3.1 (1:1,000; Alomone Labs), Kir3.4 (1:1,000; Alomone Labs), calnexin (1:2,000; Nordic Biosite), or Na + /K+-ATPase α1 (1:500; Santa Cruz Biotechnology) overnight.

Techniques: Expressing

Journal: The Journal of Biological Chemistry

Article Title: Histamine Induces ATP Release from Human Subcutaneous Fibroblasts, via Pannexin-1 Hemichannels, Leading to Ca 2+ Mobilization and Cell Proliferation *

doi: 10.1074/jbc.M113.460865

Figure Lengend Snippet: Histamine-induced [Ca2+]i mobilization depends on pannexin-1 hemichannel activity in cultured human subcutaneous fibroblasts. The effect of Hist (100 μm) was tested in the presence of inhibitors of connexin- and/or pannexin-containing hemichannels, namely MFQ (3 μm) (A, i), 2-octanol (1 mm) (A, ii), CBX (300 μm) (A, iii), 10Panx (100 μm) (A, iv), probenecid (150 μm) (A, v), and H1152 (3 μm) (A, vi) as well as upon blocking vesicle exocytosis with BFA (20 μm) (C). Cells were preincubated with the fluorescent calcium indicator Fluo-4 NW (see “Experimental Procedures”); [Ca2+]i transients were calibrated to the maximal calcium load produced by ionomycin (5 μm; 100% response). Black arrows, time of drug application. None of the inhibitors significantly changed base-line fluorescence when applied alone. Each point represents pooled data from n experiments. Vertical bars, S.E., shown when they exceed the symbols in size. *, p < 0.05, significant differences from Hist (100 μm) alone. B, representative confocal micrographs and blots of Cx43 and Panx1 hemichannel immunoreactivity in cultured human subcutaneous fibroblasts from several distinct individuals (a–g). β-Tubulin was used as a reference protein. Image scale bars, 60 μm.

Article Snippet: Membranes were subsequently incubated with goat anti-human P2Y 1 (1:200) (Santa Cruz Biotechnology, Inc.), rabbit anti-human P2Y 12 (1:200) (Alomone), rabbit anti-human P2Y 13 (1:200) (Alomone), rabbit anti-human Panx1 (1:250) (Novex, Invitrogen), and rabbit anti-human Cx43 (1:6000) (Abcam) in the above blocking buffer overnight at 4 °C.

Techniques: Activity Assay, Cell Culture, Blocking Assay, Produced, Fluorescence

Journal: The Journal of Biological Chemistry

Article Title: Histamine Induces ATP Release from Human Subcutaneous Fibroblasts, via Pannexin-1 Hemichannels, Leading to Ca 2+ Mobilization and Cell Proliferation *

doi: 10.1074/jbc.M113.460865

Figure Lengend Snippet: Histamine-induced single-cell [Ca2+]i oscillations and To-Pro3 dye uptake by cultured fibroblasts of the human subcutaneous tissue evaluated by time-lapse fluorescence confocal microscopy. A, fluorescence variation in cells preloaded with the fluorescent calcium indicator Fluo-4 NW (see “Experimental Procedures”); To-Pro3 was added to the superfusion fluid 6 min before Hist (100 μm) (dashed vertical line). Results are in arbitrary units (a.u.) of fluorescence; individual [Ca2+]i transients were calibrated to the corresponding maximal calcium load produced by ionomycin (5 μm; 100% response) in the same cell. B, bars represent the effect of Hist (100 μm) in the absence and in the presence of pannexin-1 hemichannel permeability blockers, CBX (300 μm) and H1152 (3 μm). Vertical bars, S.E. from n experiments. *, p < 0.05, significant differences from control values obtained in the absence of tested drugs; **, p < 0.05, significant differences compared with the effect of Hist (100 μm) alone.

Article Snippet: Membranes were subsequently incubated with goat anti-human P2Y 1 (1:200) (Santa Cruz Biotechnology, Inc.), rabbit anti-human P2Y 12 (1:200) (Alomone), rabbit anti-human P2Y 13 (1:200) (Alomone), rabbit anti-human Panx1 (1:250) (Novex, Invitrogen), and rabbit anti-human Cx43 (1:6000) (Abcam) in the above blocking buffer overnight at 4 °C.

Techniques: Cell Culture, Fluorescence, Confocal Microscopy, Produced, Permeability