Journal: International Journal of Molecular Sciences

Article Title: Improved Generation of Human Induced Pluripotent Stem Cell-Derived Cardiac Pacemaker Cells Using Novel Differentiation Protocols

doi: 10.3390/ijms23137318

Figure Lengend Snippet: Overview of the used TaqMan probes and primers.

Article Snippet: Subsequent primary antibodies were used: anti-HCN1 (IgG, rabbit, Alomone Labs, Ltd., Jerusalem, Israel), anti-HCN2 (IgG, rabbit, Alomone Labs, Ltd.), anti-HCN4 (IgG, rabbit, Alomone Labs, Ltd.), anti-NCX1 (IgG, rabbit, Alomone Labs, Ltd.), anti-Ca v 1.2 (IgG, mouse, Abcam, Cambridge, UK), anti-Na v 1.5 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx40 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx43 (IgG, rabbit, Alomone Labs, Ltd.), anti-Cx45 (IgG, mouse, Abcam) and anti- Tbx3 (IgG, mouse, Abcam), anti- Tbx18 (IgG, mouse, Santa Cruz Biotechnology Inc., Dallas, TX, USA), anti-Shox2 (IgG, mouse, Abcam) and anti-Nkx2.5 (IgG, rabbit, Abcam).

Techniques:

Journal: The Journal of comparative neurology

Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

doi: 10.1002/cne.24699

Figure Lengend Snippet: Primary antibodies

Article Snippet: All primary antibodies are listed in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Molecular marker Immunogen Host Dilution Clone Source RRID Albumin whole mouse albumin goat 1:800 poly Bethyl, A90–234A RRID:AB_67122 claudin5 clone 4C3C2, Alexa488 Synthetic peptide from the mouse claudin5 mouse 1:10000 mono Invitrogen, 352588 RRID:AB_2532189 Connexin 43 (Cx43s) PSSRASSRASSRPRPDDLEI rabbit 1:2000 poly Sigma, C6219 RRID:AB_476857 Connexin 43 (Cx43a) (C)HAQPFDFPDDNQNSK rabbit 1:10000 poly Alomone, ACC-201 RRID:AB_10917597 Connexin 40 (C)GHRFPQGYHSDKR rabbit 1:3000 poly Alomone, ACC-205 RRID:AB_2340916 Connexin 37 (C)EHQMAKISVAEDGR rabbit 1:2000 poly Alomone, ACC-204 RRID:AB_2722598 Smooth muscle actin, SMA, 1A4 N-terminal synthetic decapeptide of α-SMA mouse 1:1000 mono Sigma, A5228 RRID:AB_262054 Glial fibrillary acidic protein, GFAP purified bovine GFAP chicken 1:3000 poly Millipore, AB5541 RRID:AB_177521 CD31/PECAM-1 Mouse myeloma cell NS0-derived recombinant mouse CD31 Glu18-Lys590 goat 1:6000 poly RD, AF3628 RRID:AB_2161028 Open in a separate window Primary antibodies

Techniques: Marker, Purification, Recombinant

Journal: The Journal of comparative neurology

Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

doi: 10.1002/cne.24699

Figure Lengend Snippet: Primary antibodies

Article Snippet: Connexin 40 , (C)GHRFPQGYHSDKR , rabbit , 1:3000 , poly , Alomone, ACC-205 , RRID:AB_2340916.

Techniques: Marker, Purification, Recombinant

Journal: The Journal of comparative neurology

Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

doi: 10.1002/cne.24699

Figure Lengend Snippet: (a–d) Cx37 labeling revealed just a few puncta (green) on blood vessels (blue) in pure PFA fixative (a) and entire outlines of contractile cells in carbodiimide-based fixatives (b and c). Ganglion cells were more visible in PFA fixative (asterisks); the lowest background was in carbodiimide. Pre-absorption of the antibody with a control peptide eliminated the labeling (d). (e–h) Cx40 (green) was associated with outlines of endothelial cells (blue) in all fixative with the less background in carbodiimide (f). Incubation of primary antibody eliminated the labeling (h) with the exception of labeled microglia (arrow). (i–k) Cx43s-positive puncta (green, Sigma antibody) were associated with vasculature in PFA fixative (i); in addition string-like structures along endothelial cell contacts (blue) became visible in carbodiimide fixative (j–k). (l) Omission of the primary antibody eliminated punctate and string-like structures; the microglia was labeled by the secondary antibody (arrow). (m–p) Cx43a produced identical pattern as Cx43s in carbodiimide containing fixatives (n, o). In pure PFA fixative, this antibody labeled well punctate and string-like structures; however the background was significantly higher (m). Incubation of the primary antibodies with control peptide eliminated the specific labeling (p). (q–s) Morphology of DsRed expressing pericytes was best preserved in PFA-containing fixatives (q and s); the DsRed labeling in pericyte processes was lost in pure carbodiimide fixative (q). (t) Summary of fixation procedure. Red crosses indicate signal, black crosses – background; the number of crosses indicates intensity. Scale bar 10 μm. Cx43s – Cx43 antibody from Sigma, Cx43a – Cx43 antibody from Alomone, NB – Neurobiotin.

Article Snippet: Connexin 40 , (C)GHRFPQGYHSDKR , rabbit , 1:3000 , poly , Alomone, ACC-205 , RRID:AB_2340916.

Techniques: Labeling, Incubation, Produced, Expressing

Journal: The Journal of comparative neurology

Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

doi: 10.1002/cne.24699

Figure Lengend Snippet: (a) Cx40 was associated with primary arteries and some neurons in the ganglion cell layer of the retina whole mount. Insert shows primary arteries labeled for smooth muscle actin (SMA). Scale bar 200 μm. (b) In a high magnification of the superficial vascular layer Cx40 was localized directly to arteries. Scale bar 50 μm. (c) Cx40 was not detected in neither intermediate layer nor deep layer capillaries. Scale bar 50 μm. (d) Schematic presentation of Cx40 distribution in the superficial layer. (e) Quantification of Cx40 distribution on blood vessels of the superficial layer. (f) Vertical view of the confocal z-stack through the area highlighted in (b). Scale bar 25 μm. (g) Density of Cx40 in vascular layers. (h–j) Cx40 was colocalized on endothelial cells of the primary arteries but not on the capillaries (arrow). (i) Neurobiotin injected into a smooth muscle cell did not propagate to neighboring vascular cells. Please note that the targeted SMA-positive cell constricted during Neurobiotin injection. Scale bar 25 μm.

Article Snippet: Connexin 40 , (C)GHRFPQGYHSDKR , rabbit , 1:3000 , poly , Alomone, ACC-205 , RRID:AB_2340916.

Techniques: Labeling, Injection

Journal: The Journal of comparative neurology

Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

doi: 10.1002/cne.24699

Figure Lengend Snippet: (a) Cx37 expression shown in a vertical view, created by rotation of a z-stack from the corresponding retinal whole mount region in (b). (b) Confocal projection of a retinal whole mount at the GCL. Some ganglion cells expressed Cx37 at their cell membrane (asterisks, insert). (c) Blood vessels had higher expression of Cx37 (blue) than ganglion cells (red). (d–e) Cx40 was weakly expressed in numerous nuclei of inner retinal neurons. High expression of Cx40 was detected in blood vessels (f, blue). (g–i) Cx43 was expressed in nuclei (insert) and processes of inner retinal neurons (g, four bright bands in the inner plexiform layer). The expression in neurons was lower than in blood vessels (i). Scale bar 20 μm. ONL – outer nuclear layer, OPL – outer plexiform layer, INL – inner nuclear layer, IPL – inner plexiform layer, GCL – ganglion cell layer.

Article Snippet: Connexin 40 , (C)GHRFPQGYHSDKR , rabbit , 1:3000 , poly , Alomone, ACC-205 , RRID:AB_2340916.

Techniques: Expressing

Journal: The Journal of comparative neurology

Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

doi: 10.1002/cne.24699

Figure Lengend Snippet: (a–d) In the artery, Cx40 (green) was strictly colocalized with tight junctions (magenta, arrows) and less precisely with adherens proteins (blue). In a merged image of claudin5 and PECAM (d), the colocalization was not precise. White line highlights the region used for fluorescent intensity profile. Note that claudin5 and Cx40 profiles closely match and differ from the PECAM profile. (e–h) In the vascular relay, Cx43-positive strings were colocalized with tight junctions including strings and small varicosities (arrows). (i–l) In the capillary, Cx43 was not detected along tight junctions. Scale bar 10 μm.

Article Snippet: Connexin 40 , (C)GHRFPQGYHSDKR , rabbit , 1:3000 , poly , Alomone, ACC-205 , RRID:AB_2340916.

Techniques: