monoclonal anti cx43  (Cell Signaling Technology Inc)

Journal: MedComm

Article Title: Peroxisome Proliferator‑Activated Receptor Gamma Coactivator‑1α Deficiency in Hippocampal Astrocytes Underlies Enhanced Fear Memory Retrieval in Male Posttraumatic Stress Disorder Model Mice

doi: 10.1002/mco2.70671

Figure Lengend Snippet: Reduction of AST PGC‐1α–CX43 axis in SPS mice was responsible for the enhanced fear retrieval. (A) Experimental steps of FACS ( n = 3/group); (B) gate strategy and sorting results; (C) representative bands and quantitative data of PGC‐1α and NRF1 proteins; (D) representative bands and quantitative data of hippocampal CX43 protein in the control group and on Days 1, 7, and 14 after SPS modeling; (E) flowchart of the experiment ( n = 6/group); (F and G) freezing time of Con+CSF and Con+GAP27 group mice in the contextual fear test (F) and the cued fear test (G); (H) ELISA for detecting the ATP levels in the hippocampal tissues of the mice in the Con+CSF and Con+GAP27 groups; (I) ELISA for detecting the ATP levels in the hippocampal tissues of the mice in the control and SPS groups. Data are presented as mean and SEM. * p < 0.05, *** p < 0.001 between groups ( t ‐test or Tukey test).

Article Snippet: Gap 27, a CX43 gap junction blocker (HY‐P0139; MCE), was intracranially administered at 0.5 μL per side with a concentration of 100 μM [ ].

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: MedComm

Article Title: Peroxisome Proliferator‑Activated Receptor Gamma Coactivator‑1α Deficiency in Hippocampal Astrocytes Underlies Enhanced Fear Memory Retrieval in Male Posttraumatic Stress Disorder Model Mice

doi: 10.1002/mco2.70671

Figure Lengend Snippet: AST PGC‐1α knockdown enhanced fear memory and anxiety‐like behaviors and impaired AST structure. (A) Flowchart of the experiment ( n = 9/group); (B) representative images of virus‐labeled astrocytes; (C) representative bands and quantitative data of PGC‐1α and CX43; (D) freezing of mice in the contextual fear test; (E) freezing of mice in the cued fear test; (F) the percentage of time that mice spent exploring the open arm in the elevated maze test out of the total time; (G and H) the total distance and representative traces of the open field test; (I and J) the results and representative images of the Sholl analysis of astrocytes. The data are presented as mean ± standard error ( n = 3/group). (K) Representative image of S100A10 immunofluorescence staining (scale bar: 50 µm); (L) the percentage of colabeling of S100A10 and PGC‐1α ( n = 3/group). Data are presented as mean and SEM. *p < 0.05, **p < 0.01, ***p < 0.001 between groups ( t ‐test).

Article Snippet: Gap 27, a CX43 gap junction blocker (HY‐P0139; MCE), was intracranially administered at 0.5 μL per side with a concentration of 100 μM [ ].

Techniques: Knockdown, Virus, Labeling, Immunofluorescence, Staining

Journal: MedComm

Article Title: Peroxisome Proliferator‑Activated Receptor Gamma Coactivator‑1α Deficiency in Hippocampal Astrocytes Underlies Enhanced Fear Memory Retrieval in Male Posttraumatic Stress Disorder Model Mice

doi: 10.1002/mco2.70671

Figure Lengend Snippet: Activation of hippocampal AST or PGC‐1α reduced fear memory retrieval in SPS mice. (A) Flowchart of experiment 5 ( n = 7/group); (B) representative image of immunofluorescence of hM3Dq‐labeled AST; (C) freezing time of mice in the contextual fear test; (D) freezing time of mice in the cued fear test; (E) percentage of time that mice spent exploring the open arm in the elevated maze test out of the total time; (F) flowchart of the experiment 5 ( n = 6/group); (G) freezing time of mice in the contextual fear memory; (H) freezing time of mice in the cued fear memory; (I) percentage of time that mice spent exploring the open arm in the elevated maze test out of the total time; (J and K) representative bands of GFAP (J) and CX43 (K) along with quantitative data; (L) flowchart of the experiment 6 ( n = 7/group); (M) schematic diagram of the hM3Dq–mCherry/AAV5–PGC‐1α viral injection site and colabeling; (N) freezing time of mice in the contextual fear memory; (O) freezing time of mice in the cued fear memory. Data are presented as mean and SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 between groups ( t ‐test or Tukey test).

Article Snippet: Gap 27, a CX43 gap junction blocker (HY‐P0139; MCE), was intracranially administered at 0.5 μL per side with a concentration of 100 μM [ ].

Techniques: Activation Assay, Immunofluorescence, Labeling, Injection

Journal: Science Advances

Article Title: In situ structure of the human gap junction

doi: 10.1126/sciadv.aea4183

Figure Lengend Snippet: ( A ) A schematic representation of the gap junction assembly from connexin 43 (Cx) monomers to hemichannels and to the gap junction plaque between two cells. Intra- (IL) and extracellular (EL) loops are labeled. The transmembrane (TM) helices are numbered 1 to 4. ( B ) One-nm-thick slice through a cryo-ET reconstruction of a cell-cell junction. Areas harboring Cx43 channels (green), 80 S ribosomes (cyan), and microtubules (pink) have been indicated. A mitochondrion has been labeled (M). Scale bars, 100 nm. ( C ) Close-up of the area indicated in (B). Side views of Cx43 GJCs bridging two plasma membranes are indicated with arrowheads. Scale bar, 10 nm. ( D ) Close-up from a different cryo-ET reconstruction showing top views of the Cx43 GJCs. Scale bar, 10 nm. ( E ) Results of template matching for Cx43 GJCs, 80 S ribosomes, and microtubule segments are shown after plotting the templates back onto the cryo-ET tomogram (illustrated with a 1-nm-thick slice in the background). The gap junction is depicted as a transparent surface (green) fitted to the Cx43 channel positions for visual clarity. ( F ) Histogram of pairwise distances between 80 S ribosomes and microtubule segments (MTs) to the Cx43 GJCs, calculated from multiple cryo-ET reconstructions ( n = 26). The 15-nm-wide zone occupied by Cx43 channels is indicated in green. The 22-nm-wide ribosome exclusion zone (REZ) is indicated in gray.

Article Snippet: The cryo-ET map of the Cx43 lattice has been deposited in the Electron Microscopy DataBank (EMDB) with the deposition ID EMD-54024.

Techniques: Labeling, Tomography, Clinical Proteomics

Journal: Science Advances

Article Title: In situ structure of the human gap junction

doi: 10.1126/sciadv.aea4183

Figure Lengend Snippet: ( A ) Slice through a subtomogram average of the Cx43 lattice is shown. One channel is indicated (arrowhead). Scale bar, 10 nm for (A) to (C). ( B ) Slice through the subtomogram average, along the solid line in (A). ( C ) Slice along the dashed line in (A). Channels are indicated in (B) and (C) (arrowheads). ( D ) Isosurface of a subtomogram average. A low-pass filter to 20-Å resolution has been applied to improve the interpretability of the lateral layers. Cx43 atomic models (PDB: 7Z22, residues 17 to 105 and 151 to 235) are shown in green. Two hemichannels forming a full channel are indicated (arrowheads). The convex (+) and concave (−) sides of the lattice are indicated in (B) to (D). ( E ) Density distribution as a function of distance from the lattice midpoint is shown. Density is in arbitrary units. The inner (IL) and outer (OL) leaflets of the two lipid bilayers are marked. The regions corresponding to intracellular density bridging Cx43 hexamers, denoted as lateral-contact layers (LCLs), are labeled. An additional layer of density on the concave side of the gap junction is indicated with an asterisk. ( F to H ) Isosurface renderings are shown from the top (+ to – direction) for the extracellular region (gap), inner leaflet (IL1), and intracellular densities (OL1). ( I ) Rendering of the CG MD simulation setup. The connexins are green, the POPC lipids are blue (head groups in a darker shade), and cholesterol is magenta. The solvent water is rendered as a transparent cube. The inset shows the area indicated. ( J and K ) Number of POPC lipids (J) and cholesterol molecules (K) around the centermost gap junction hemichannel is plotted as a function of the simulation time for both the membranes (+ and – sides).

Article Snippet: The cryo-ET map of the Cx43 lattice has been deposited in the Electron Microscopy DataBank (EMDB) with the deposition ID EMD-54024.

Techniques: Labeling, Solvent

Journal: Science Advances

Article Title: In situ structure of the human gap junction

doi: 10.1126/sciadv.aea4183

Figure Lengend Snippet: ( A to C ) Isosurface renderings of the cryo-ET subtomogram average at 14-Å resolution segmented with a cylindrical mask are shown as gray transparent surfaces, together with a partial atomic model of the Cx43 channel (PDB: 7Z22, residues 17 to 105 and 151 to 235). The inset indicates the level of the cross sections taken from the middle of the top bilayer (A), the middle of the extracellular loops (B), and the middle of the bottom bilayer (C). The TM helices are numbered 1 to 4 for one Cx monomer in both hemichannels. The convex (+) and concave (−) sides of the gap junction are indicated. ( D ) Close-up of the hemichannel cryo-ET map (this study) and a single-particle cryo-EM structure of the Cx43 hemichannel (EMD-14475), low-pass filtered to the same resolution (14 Å) for comparison. At this resolution, the N-terminal helices (residues 1 to 16, purple) closing the channel are visible in the cryo-EM density. The dashed lines indicate the position of the two membrane leaflets in both structures. ( E and F ) Close-ups of the intracellular areas indicated in the inset (dashed rectangles) are shown. In both, a stalk-like density can be seen, connecting the density below it (stem) and the density above it [lateral contacts layer (LCL)].

Article Snippet: The cryo-ET map of the Cx43 lattice has been deposited in the Electron Microscopy DataBank (EMDB) with the deposition ID EMD-54024.

Techniques: Tomography, Single Particle, Cryo-EM Sample Prep, Comparison, Membrane

Journal: European Heart Journal

Article Title: Left bundle branch vs biventricular pacing: mechanistic insights from a canine model

doi: 10.1093/eurheartj/ehaf1093

Figure Lengend Snippet: Molecular targets in cardiac remodeling by BiVP and LBBP. Western blotting results of BNP, TGF-β, KCNIP2, SERCA2a, Cx43, ANKRD1 protein expression (A, C, E) in septum and LV lateral walls from NC, DHF, and BiVP and LBBP groups, and relative protein expression (B, D, F). Represented images and statistical data (G) of cardiac fibrosis detected using Masson's staining. † P < .05 between septal and lateral walls. Abbreviations as in

Article Snippet: The levels of several key proteins were examined using western blot, including ANKRD1 (Proteintech, 11427-1-AP), BNP (Abclonal, A23996), Cx43 (Proteintech, 26980-1-AP), GSNOR (Proteintech, 66193-1-Ig), GLUD1 (Proteintech, 14299-1-AP), KCNIP2 (Abclonal, A7100), SERCA2a (Abclonal, A0098), TGF-β (Proteintech, 81746-2-RR), and GAPDH (Proteintech, 10494-1-AP).

Techniques: Western Blot, Expressing, Staining

Journal: European Heart Journal

Article Title: Left bundle branch vs biventricular pacing: mechanistic insights from a canine model

doi: 10.1093/eurheartj/ehaf1093

Figure Lengend Snippet: Overview of the key findings. This study compares the treatment efficacy of LBBP and BiVP, and relating electrocardiographic, echocardiographic and cellular (reverse) remodeling in a canine DHF model. The results show that LBBP restores electrical synchrony (QRSd) to a similar extent as BiVP, while leads to larger LV-GLS improvement than BiVP. Both LBBP and BiVP reverse the protein level of KCNIP2, SERCA2a and Cx43. Moreover, LBBP reverses abnormalities in cytoskeleton-associated proteins, TGF-β signaling, and SERCA2a expression more than BiVP, as well as enhances myocardial energy metabolism via more efficient TCA cycling and ATP production. ANKRD1, ankyrin repeat domain 1; ATP, adenosine triphosphate; BiVP, biventricular pacing; BNP, B-type natriuretic peptide; Cx43, connexin 43; DHF dyssynchronous heart failure; ECG, electrocardiogram; echo, echocardiogram; GLUD1, glutamate dehydrogenase 1; GSNOR S-nitrosoglutathione reductase; KCNIP2, potassium voltage-gated channel interacting protein 2; LBBP, left bundle branch pacing; LV-GLS, left ventricular global longitudinal strain; NC, normal control; SERCA2a, sarcoplasmic/endoplasmic reticulum Ca 2+ ATPase 2a; TCA, tricarboxylic acid; TGF-β, Transforming growth factor-β.

Article Snippet: The levels of several key proteins were examined using western blot, including ANKRD1 (Proteintech, 11427-1-AP), BNP (Abclonal, A23996), Cx43 (Proteintech, 26980-1-AP), GSNOR (Proteintech, 66193-1-Ig), GLUD1 (Proteintech, 14299-1-AP), KCNIP2 (Abclonal, A7100), SERCA2a (Abclonal, A0098), TGF-β (Proteintech, 81746-2-RR), and GAPDH (Proteintech, 10494-1-AP).

Techniques: Expressing, Control