Journal: The FASEB Journal

Article Title: Connexin 43 gap junctions contribute to brain endothelial barrier hyperpermeability in familial cerebral cavernous malformations type III by modulating tight junction structure

doi: 10.1096/fj.201700699R

Figure Lengend Snippet: Depletion of Cx43 protein and GAP27-mediated inhibition of Cx43 GJIC rescues CCM3KD monolayer permeability. A) Transwell TEER assays revealed a significant increase in CCM3KD permeability and partial, significant rescue after the reduction of Cx43 expression in CCM3KD cells via Cx43 siRNA-mediated KD. Data represent means ± sem (n = 3–10 independent experiments). *P < 0.05, ***P < 0.0001 compared with control, ***P < 0.001 compared with CCM3KD. Cx43 + CCM3KD efficiency is reported in Supplemental Fig. 2. B) Permeability coefficient (PC) of inulin (MW = 5 kDa) shows increased permeability of CCM3KD and Cx43over mBEC monolayers, whereas the depletion of Cx43 via siRNA rescued mBEC monolayer integrity. Data represent means ± sem. ***P < 0.001 compared with control, ***P < 0.001 compared with CCM3KD. C) Transwell TEER assays performed with control and CCM3KD cells treated with 100 μm GAP19 or 100 μM GAP27 for 24 h. Treatment with GAP27 completely rescued CCM3KD permeability. GAP19 treatment had no effect (n = 3–10 independent experiments). ***P < 0.0001 compared with control or CCM3KD. D) Transendothelial albumin permeability was only slightly increased in CCM3KD and Cx43over monolayers compared with control. Inhibition of vesicular transport with N-ethylmaleimide (NEM) did not disturb the albumin monolayer permeability, which indicated that CCM3 and Cx43 predominantly regulate paracellular permeability. Data represent means ± sem. *P < 0.05 compared with control.

Article Snippet: Inhibition studies Inhibition of Cx43 GJs and HCs was achieved by cell treatment with 100 μM GAP27 (Cx43 gap junction and hemichannel inhibitor) or 100 μM GAP19 (Cx43 hemichannel inhibitor; Tocris Biosciences, Bristol, United Kingdom), respectively, for 24 h. Treatment did not affect cell viability as tested by Live Dead assay (Thermo Fisher Scientific).

Techniques: Inhibition, Permeability, Expressing, Control

Journal: IBRO Neuroscience Reports

Article Title: Acupuncture regulates astrocyte neurotoxic polarization to protect blood–brain barrier integrity in delayed thrombolysis through mediating ERK1/2/Cx43 axis

doi: 10.1016/j.ibneur.2025.04.005

Figure Lengend Snippet: Cx43 inhibition reduces neurotoxic polarization of astrocytes and inflammatory responses. (A) Representative IF images showed co-localization of Cx43 (red) and GFAP (Green) in the Sham, rt-PA, and Tat-Gap19-treated groups. (B) Quantification of GFAP + /Cx43 + intensities in the brain cortex of ischemic penumbra from different groups (n = 3, Scale bar=20μm). (C) Western blot analysis of C3 expression in the groups. (D) Quantification of C3 protein expression in the western blot analysis (n = 3). (E) Double immunofluorescence staining for C3 (red) and GFAP (green) in the ischemic penumbra. (F) the number of C3 + /GFAP + cells were quantified in each 1 mm 2 area of the ischemic penumbra (n = 3, Scale bar=20μm). (G-I) Secretion of inflammatory cytokines IL-1β, IL-6 and TNF-α (n = 5). Data was presented as mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The injection protocol for Tat-Gap19 and Ceramide C6 was as follows: Tat-Gap19 (1 μg/μl, MCE, USA) and Ceramide C6 (0.5 μg/μl, SANTA, USA) were injected at a volume of 5 μl each at coordinates 1.0 mm posterior and 1.5 mm lateral from the bregma, with a depth of 4 mm.

Techniques: Inhibition, Western Blot, Expressing, Double Immunofluorescence Staining

Journal: Pharmaceuticals

Article Title: Effects of an Atypical Antipsychotic, Zotepine, on Astroglial L-Glutamate Release through Hemichannels: Exploring the Mechanism of Mood-Stabilising Antipsychotic Actions and Antipsychotic-Induced Convulsion

doi: 10.3390/ph14111116

Figure Lengend Snippet: Effects of selective Cx43 inhibitor, N-terminal transactivator of transcription GAP19 (TAT-GAP19) (10 μM) on stimulatory effects of supratherapeutic concentration of ZTP (1000 nM) on basal ( A ) and HK-ACSF-evoked ( B ) astroglial L-glutamate releases. Ordinate: mean ± SD (n = 6) of astroglial L-glutamate release (μM). Light and dark colour columns indicate the incubation without (non) and with TAT-GAP19, respectively. ** p < 0.01: relative to non, @ p < 0.05, @@ p < 0.01 relative to control by MANOVA with Tukey’s (wholly significant difference) post-hoc test.

Article Snippet: Zotepine (ZTP), selective Cx43 inhibitor, N-terminal transactivator of transcription GAP19 (TAT-GAP19) and Akt inhibitor, 10-[4’-(N,N-diethylamino)butyl]-2-chlorophenoxazine hydrochloride (DEBC) were obtained from Funakoshi (Tokyo, Japan).

Techniques: Concentration Assay, Incubation, Control

Journal: Pharmaceuticals

Article Title: Effects of an Atypical Antipsychotic, Zotepine, on Astroglial L-Glutamate Release through Hemichannels: Exploring the Mechanism of Mood-Stabilising Antipsychotic Actions and Antipsychotic-Induced Convulsion

doi: 10.3390/ph14111116

Figure Lengend Snippet: Effects of acute (120 min) administration of supratherapeutic concentration of ZTP (1000 nM) and chronic (14 days) administration of therapeutically relevant concentrations of ZTP (300 nM) on Cx43 protein expression in the plasma membrane fraction ( A ) and their pseudogel images, using capillary immunoblotting ( B ). Ordinate: mean ± SD (n = 6) of the relative protein level of Cx43 (per GAPDH). Effects of ZTP on Cx43 expression in the plasma membrane fraction of the primary cultured astrocytes were analysed by student T-test (* p < 0.05 vs. control: ZTP free).

Article Snippet: Zotepine (ZTP), selective Cx43 inhibitor, N-terminal transactivator of transcription GAP19 (TAT-GAP19) and Akt inhibitor, 10-[4’-(N,N-diethylamino)butyl]-2-chlorophenoxazine hydrochloride (DEBC) were obtained from Funakoshi (Tokyo, Japan).

Techniques: Concentration Assay, Expressing, Clinical Proteomics, Membrane, Western Blot, Cell Culture, Control

Journal: Pharmaceuticals

Article Title: Effects of an Atypical Antipsychotic, Zotepine, on Astroglial L-Glutamate Release through Hemichannels: Exploring the Mechanism of Mood-Stabilising Antipsychotic Actions and Antipsychotic-Induced Convulsion

doi: 10.3390/ph14111116

Figure Lengend Snippet: Concentration-dependent effects of subchronic administration of ZTP on Cx43 protein expression in the cytosol fraction ( A ) and their pseudogel images, using capillary immunoblotting ( B ). Ordinate: mean ± SD (n = 6) of the relative protein level of Cx43 (per GAPDH). Concentration-dependent effects of ZTP (100, 300 and 1000 nM) on Cx43 expression in the cytosol fraction of the primary cultured astrocytes were analysed by one-way ANOVA with Tukey’s (wholly significant difference) post hoc test (* p < 0.05 vs. ZTP free (0)).

Article Snippet: Zotepine (ZTP), selective Cx43 inhibitor, N-terminal transactivator of transcription GAP19 (TAT-GAP19) and Akt inhibitor, 10-[4’-(N,N-diethylamino)butyl]-2-chlorophenoxazine hydrochloride (DEBC) were obtained from Funakoshi (Tokyo, Japan).

Techniques: Concentration Assay, Expressing, Western Blot, Cell Culture

Journal: Pharmaceuticals

Article Title: Effects of an Atypical Antipsychotic, Zotepine, on Astroglial L-Glutamate Release through Hemichannels: Exploring the Mechanism of Mood-Stabilising Antipsychotic Actions and Antipsychotic-Induced Convulsion

doi: 10.3390/ph14111116

Figure Lengend Snippet: Concentration-dependent effects of subchronic administration of ZTP on Cx43 protein expression in the plasma membrane fraction ( A ) and their pseudogel images, using capillary immunoblotting ( B ). Ordinate: mean ± SD (n = 6) of the relative protein level of Cx43 (per GAPDH). Concentration-dependent effects of ZTP (100, 300 and 1000 nM) on Cx43 expression in the plasma membrane fraction of the primary cultured astrocytes were analysed by one-way ANOVA with Tukey’s (wholly significant difference) post hoc test (** p < 0.01 vs. ZTP free (0)).

Article Snippet: Zotepine (ZTP), selective Cx43 inhibitor, N-terminal transactivator of transcription GAP19 (TAT-GAP19) and Akt inhibitor, 10-[4’-(N,N-diethylamino)butyl]-2-chlorophenoxazine hydrochloride (DEBC) were obtained from Funakoshi (Tokyo, Japan).

Techniques: Concentration Assay, Expressing, Clinical Proteomics, Membrane, Western Blot, Cell Culture

Journal: Pharmaceuticals

Article Title: Effects of an Atypical Antipsychotic, Zotepine, on Astroglial L-Glutamate Release through Hemichannels: Exploring the Mechanism of Mood-Stabilising Antipsychotic Actions and Antipsychotic-Induced Convulsion

doi: 10.3390/ph14111116

Figure Lengend Snippet: Interaction between subchronic administration of supratherapeutic concentration of ZTP and Akt inhibitor (DEBC) on Cx43 protein expression in the astroglial plasma membrane fraction ( A ) and their pseudogel images, using capillary immunoblotting ( B ). Ordinate: mean ± SD (n = 6) of the relative protein level of Cx43 (per GAPDH). Effects of ZTP (1000 nM) and Akt inhibitor (DEBC: 10 μM) on Cx43 expression in the plasma membrane fraction of the primary cultured astrocytes were analysed by two-way ANOVA with Tukey’s (wholly significant difference) post hoc test (** p < 0.01 vs. control, @@ p < 0.01 vs. non).

Article Snippet: Zotepine (ZTP), selective Cx43 inhibitor, N-terminal transactivator of transcription GAP19 (TAT-GAP19) and Akt inhibitor, 10-[4’-(N,N-diethylamino)butyl]-2-chlorophenoxazine hydrochloride (DEBC) were obtained from Funakoshi (Tokyo, Japan).

Techniques: Concentration Assay, Expressing, Clinical Proteomics, Membrane, Western Blot, Cell Culture, Control

Journal: Pharmaceuticals

Article Title: Effects of an Atypical Antipsychotic, Zotepine, on Astroglial L-Glutamate Release through Hemichannels: Exploring the Mechanism of Mood-Stabilising Antipsychotic Actions and Antipsychotic-Induced Convulsion

doi: 10.3390/ph14111116

Figure Lengend Snippet: Summary of the effects of zotepine (ZTP) on astroglial L-glutamate release and Cx43 expression in the astrocyte.

Article Snippet: Zotepine (ZTP), selective Cx43 inhibitor, N-terminal transactivator of transcription GAP19 (TAT-GAP19) and Akt inhibitor, 10-[4’-(N,N-diethylamino)butyl]-2-chlorophenoxazine hydrochloride (DEBC) were obtained from Funakoshi (Tokyo, Japan).

Techniques: Expressing, Clinical Proteomics, Membrane