Journal:

Article Title: Modulation of Astrocyte P2Y 1 Receptors by the Carboxyl Terminal Domain of the Gap Junction Protein Cx43

doi: 10.1002/glia.20598

Figure Lengend Snippet: (A) Western blot showing expression of Cx43 in WT (lane 1) and in Cx43-null astrocytes transfected with full length Cx43 (lane 3), with Cx43 carboxyl terminus (lane 4) and with Cx43M257 (lane 5). Lane 2 corresponds to untransfected Cx43-null cells. Lanes 1–4 correspond to immunoblots performed with Cx43-18A antibody and lane 5 with Cx43-16A. (B) Time courses of calcein FRAP obtained from untreated (black squares) and 48 h carbenoxolone-treated (open triangles) WT astrocytes, and from untransfected (open circles) and transfected Cx43-null astrocytes with full length Cx43 (open squares) and Cx43M257 (black circles). Data were obtained from astrocytes cultured from a minimum of 3 litters of mice.

Article Snippet: The following primary antibodies were employed: polyclonal antiP2Y1 (1:200; Alomone Labs), and two different polyclonal antiCx43 antibodies (gifts from Dr. E.L. Hertzberg, Albert Einstein College of Medicine), one recognizing Cx43 at position 241–261 (16A) and the other at position 345–360 (18A).

Techniques: Western Blot, Expressing, Transfection, Cell Culture

Journal:

Article Title: Modulation of Astrocyte P2Y 1 Receptors by the Carboxyl Terminal Domain of the Gap Junction Protein Cx43

doi: 10.1002/glia.20598

Figure Lengend Snippet: (A) Dose-response curves obtained for untreated (black squares) and 48 h carbenoxolone-treated (open triangles) WT and untransfected Cx43-null spinal cord astrocytes exposed to the P2Y1R agonist 2-MeS-ATP. Note that blockade of gap junctional communication in WT astrocytes does not alter the half-maximal response (EC50 value) induced by the P2Y1R agonist. Mean values were obtained from four to seven independent experiments. (B) Dose-response curves obtained for WT (black squares), untransfected (open circles) and transfected Cx43-null astrocytes with full length Cx43 (open squares), Cx43M257 (black circles) and Cx43CT (black triangles). Note that both full length Cx43 and Cx43CT but not Cx43M257 shifted the EC50 values of 2-MeS-ATP obtained for Cx43-null astrocytes to those obtained in WT cells. Data were obtained from 5 to 8 litters.

Article Snippet: The following primary antibodies were employed: polyclonal antiP2Y1 (1:200; Alomone Labs), and two different polyclonal antiCx43 antibodies (gifts from Dr. E.L. Hertzberg, Albert Einstein College of Medicine), one recognizing Cx43 at position 241–261 (16A) and the other at position 345–360 (18A).

Techniques: Transfection

Journal:

Article Title: Modulation of Astrocyte P2Y 1 Receptors by the Carboxyl Terminal Domain of the Gap Junction Protein Cx43

doi: 10.1002/glia.20598

Figure Lengend Snippet: Bar histograms showing the mean values of P2Y1R expression levels in WT, untransfected Cx43-null (KO), and in Cx43-null transfected with Cx43 CT, Cx43 truncated at position 257 (M257) and with full length Cx43. An example of western blots for Cx43 and β-actin is shown above.

Article Snippet: The following primary antibodies were employed: polyclonal antiP2Y1 (1:200; Alomone Labs), and two different polyclonal antiCx43 antibodies (gifts from Dr. E.L. Hertzberg, Albert Einstein College of Medicine), one recognizing Cx43 at position 241–261 (16A) and the other at position 345–360 (18A).

Techniques: Expressing, Transfection, Western Blot

Journal:

Article Title: Modulation of Astrocyte P2Y 1 Receptors by the Carboxyl Terminal Domain of the Gap Junction Protein Cx43

doi: 10.1002/glia.20598

Figure Lengend Snippet: (A) Dose-response curves obtained for WT (black squares) and untransfected (open circles) and Cx43Δ260–280 transfected (black triangles) Cx43-null astrocytes exposed to 2-MeS-ATP, showing that deletion of the Cx43 SH3 domain does not rescue P2Y1 receptor function. Mean±SE values are from 100 to 200 cells obtained from three litters. (B) Dose-response curves obtained from WT (black squares), and Cx43-null astrocytes untreated (open circles) and treated (black triangles) with a membrane permeant peptide corresponding to amino acids 260–280 of Cx43CT. (C) Bar histograms of the mean±SE values of intracellular calcium mobilization induced by 100 nM 2-MeS-ATP recorded from WT, Cx43-null and Cx43-null transfected with Cx43CT and Cx43M257 in the absence and presence of 5 µM PP2. Note that only untransfected and Cx43M257 transfected Cx43-null astrocytes that were not exposed to PP2 did not respond to agonist with intracellular calcium levels similar to WT astrocytes. Mean ± SE values are from 4 litters (**P < 0.001; ANOVA followed by Newman-Keuls’ Multiple Comparison Test).

Article Snippet: The following primary antibodies were employed: polyclonal antiP2Y1 (1:200; Alomone Labs), and two different polyclonal antiCx43 antibodies (gifts from Dr. E.L. Hertzberg, Albert Einstein College of Medicine), one recognizing Cx43 at position 241–261 (16A) and the other at position 345–360 (18A).

Techniques: Transfection

Journal:

Article Title: Modulation of Astrocyte P2Y 1 Receptors by the Carboxyl Terminal Domain of the Gap Junction Protein Cx43

doi: 10.1002/glia.20598

Figure Lengend Snippet: (A) Western blot showing decreased expression levels of Cx43 following exposure of spinal cord astrocytes to IL-1β (B) Dose-response curves obtained for 2-MeS-ATP performed on Fura-2 loaded WT and Cx43 KO astrocytes treated for 24 h with IL-1β (20 ng/mL). Note that exposure to the cytokine altered the agonist EC50 values in WT astrocytes. About 180 cells from three independent experiments were used in each condition.

Article Snippet: The following primary antibodies were employed: polyclonal antiP2Y1 (1:200; Alomone Labs), and two different polyclonal antiCx43 antibodies (gifts from Dr. E.L. Hertzberg, Albert Einstein College of Medicine), one recognizing Cx43 at position 241–261 (16A) and the other at position 345–360 (18A).

Techniques: Western Blot, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Hypoxia-preconditioning human bone marrow-derived mesenchymal stem cells induce high-quality mitochondrial transfer through gap junctions to alleviate ischemia-reperfusion injury in liver graft

doi: 10.1186/s12964-025-02497-1

Figure Lengend Snippet: Hypo-hBMSCs enhance GJs function via up-regulating Cx43 and Cx32 expression. A Public databases indicated the protein expression levels of the Cx in hBMSCs and liver tissues. B Western blotting showed levels of Cx43, Cx32 and Cx26 protein expression in hBMSCs and BRL3A cells. GAPDH served as a loading control. C RT-qPCR and Western blotting were used to detect Cx43, Cx32 and Cx26 mRNAs and proteins levels in hBMSCs/hypo-hBMSCs. β-actin served as a loading control. D Co-IP assay was used to determine whether heterotypic channels were formed. β-actin served as a loading control. Data are shown as mean ± SEM, n = 3–5. * p < 0.05, ** p < 0.01, *** p < 0.001. HBMSCs, human bone marrow mesenchymal stem cells; hypo-hBMSCs, hypoxia preconditioning-hBMSCs

Article Snippet: To establish a HR cell model, BRL3A cells with or without hBMSCs were processed using the same protocol as previously described for the hBMSCs, and placed in a Galaxy 48R hypoxia incubator (Eppendorf GmbH, Hamburg, Germany) with 5% CO 2 and 1% O 2 at 37 °C for 12 h. Subsequently, the cells were placed in the normal cell incubator for 2 h. To plumb the role of Cx43 and Cx32 in this study, retinoic acid (RA, 1 μM, MedChemExpress, New Jersey, USA), the specific GJs agonist, and Gap26 (0.25 mg/mL, APExBIO, Houston, USA), the specific GJs inhibitor, were used in hBMSCs.

Techniques: Expressing, Western Blot, Control, Quantitative RT-PCR, Co-Immunoprecipitation Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Hypoxia-preconditioning human bone marrow-derived mesenchymal stem cells induce high-quality mitochondrial transfer through gap junctions to alleviate ischemia-reperfusion injury in liver graft

doi: 10.1186/s12964-025-02497-1

Figure Lengend Snippet: Functional homotypic Cx43-GJs and Cx32-GJs mediate mitochondrial transfer from hypo-hBMSCs to hepatocytes. A Western blotting showed levels of Cx43, Cx32 in hBMSCs. GAPDH and β-actin served as a loading control. B - E Evaluation of GJs function and mitochondrial transfer. (B, D) “Parachute” dye-coupling assay measured the function of GJs. (C, E) Immunofluorescence staining showed the levels of mitochondrial transfer from hBMSCs or hypo-hBMSCs to BRL3A cells. Data are shown as mean ± SEM, n = 3–5. * p < 0.05, ** p < 0.01, *** p < 0.001. HBMSCs, human bone marrow mesenchymal stem cells; hypo-hBMSCs, hypoxia preconditioning-hBMSCs

Article Snippet: To establish a HR cell model, BRL3A cells with or without hBMSCs were processed using the same protocol as previously described for the hBMSCs, and placed in a Galaxy 48R hypoxia incubator (Eppendorf GmbH, Hamburg, Germany) with 5% CO 2 and 1% O 2 at 37 °C for 12 h. Subsequently, the cells were placed in the normal cell incubator for 2 h. To plumb the role of Cx43 and Cx32 in this study, retinoic acid (RA, 1 μM, MedChemExpress, New Jersey, USA), the specific GJs agonist, and Gap26 (0.25 mg/mL, APExBIO, Houston, USA), the specific GJs inhibitor, were used in hBMSCs.

Techniques: Functional Assay, Western Blot, Control, Immunofluorescence, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Hypoxia-preconditioning human bone marrow-derived mesenchymal stem cells induce high-quality mitochondrial transfer through gap junctions to alleviate ischemia-reperfusion injury in liver graft

doi: 10.1186/s12964-025-02497-1

Figure Lengend Snippet: Functional homotypic Cx43-GJs and Cx32-GJs mediate mitochondrial transfer from hypo-hBMSCs to hepatocytes. A Western blotting showed levels of Cx43, Cx32 in hBMSCs. B “Parachute” dye-coupling assay measured the function of GJs. C Immunofluorescence staining showed the levels of mitochondrial transfer from hBMSCs or hypo-hBMSCs to BRL3A cells. Data are shown as mean ± SEM, n = 3–5. * p < 0.05, ** p < 0.01, *** p < 0.001. HBMSCs, human bone marrow mesenchymal stem cells; hypo-hBMSCs, hypoxia preconditioning-hBMSCs

Article Snippet: To establish a HR cell model, BRL3A cells with or without hBMSCs were processed using the same protocol as previously described for the hBMSCs, and placed in a Galaxy 48R hypoxia incubator (Eppendorf GmbH, Hamburg, Germany) with 5% CO 2 and 1% O 2 at 37 °C for 12 h. Subsequently, the cells were placed in the normal cell incubator for 2 h. To plumb the role of Cx43 and Cx32 in this study, retinoic acid (RA, 1 μM, MedChemExpress, New Jersey, USA), the specific GJs agonist, and Gap26 (0.25 mg/mL, APExBIO, Houston, USA), the specific GJs inhibitor, were used in hBMSCs.

Techniques: Functional Assay, Western Blot, Immunofluorescence, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Hypoxia-preconditioning human bone marrow-derived mesenchymal stem cells induce high-quality mitochondrial transfer through gap junctions to alleviate ischemia-reperfusion injury in liver graft

doi: 10.1186/s12964-025-02497-1

Figure Lengend Snippet: Graphical scheme of Hypo-hBMSCs induce high-quality mitochondrial transfer through GJs to alleviate hepatic IRI. ① Hypo-hBMSCs initiated mitophagy to improve mitochondrial quality. ② Hypo-hBMSCs enhanced the expression of Cx43 and Cx32. ③ Hypo-hBMSCs facilitated efficient mitochondrial transfer to hepatocytes by enhancing Cx43-GJs and Cx32-GJs function. ④ Hypo-hBMSCs transferred a greater quantity and higher quality of mitochondria into hepatocytes, enabling the hepatocytes to produce more ATP and thereby alleviating Hepatic IRI. IRI, ischemia and reperfusion injury; HR, hypoxia/reoxygenation; hBMSCs, human bone marrow mesenchymal stem cells; hypo-hBMSCs, hypoxia preconditioning-hBMSC

Article Snippet: To establish a HR cell model, BRL3A cells with or without hBMSCs were processed using the same protocol as previously described for the hBMSCs, and placed in a Galaxy 48R hypoxia incubator (Eppendorf GmbH, Hamburg, Germany) with 5% CO 2 and 1% O 2 at 37 °C for 12 h. Subsequently, the cells were placed in the normal cell incubator for 2 h. To plumb the role of Cx43 and Cx32 in this study, retinoic acid (RA, 1 μM, MedChemExpress, New Jersey, USA), the specific GJs agonist, and Gap26 (0.25 mg/mL, APExBIO, Houston, USA), the specific GJs inhibitor, were used in hBMSCs.

Techniques: Expressing

Journal: International journal of molecular medicine

Article Title: Secretome of EMSCs neutralizes LPS‑induced acute lung injury via aerosol administration.

doi: 10.3892/ijmm.2023.5307

Figure Lengend Snippet: Figure 1. Morphology and fluorescence identification of EMSCs. (A) EMSCs in P0 and P3. (B) Expression of MSC markers (CD44, Vimentin) and neural crest‑related biomarkers (Cx43, Sox9). EMSCs, ectodermal mesenchymal stem cells; Sox9, SRY‑related high‑mobility group box‑containing protein 9; Cx43, Connexin43.

Article Snippet: Briefly, samples in 24‐well plates were incubated with primary antibodies for CD44 (1:100, Boster, A00052), Cx43 (1:100, Boster, BA1727), Sox9 (1:100, Boster, PA1026‐1), Vimentin (1:100, Boster, PB9359) after being blocked with BSA.

Techniques: Fluorescence, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Cx43 Promotes Endothelial Cell Migration and Angiogenesis via the Tyrosine Phosphatase SHP-2

doi: 10.3390/ijms23010294

Figure Lengend Snippet: Downregulation of Cx43 reduces endothelial cell migration. ( a ) Western blot analysis of Cx43, Cx40, and Cx37 revealed that Cx43 is expressed in both human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC), whereas Cx40 and Cx37 are not expressed in HMEC. Cx43 (Cx43), Cx40 (Cx), and Cx37 (Cx) expressing HeLa cells were used as the positive controls. CTL: Wildtype HeLa cell lysate. ( b ) Western blot analysis of Cx43 in HMEC transfected with 100 nM siRNA against Cx43 or non-silencing control siRNA (Ctrl) for 48 h. The Cx43 expression was analysed using a polyclonal Cx43 antibody. Immunostaining for GAPDH was used as the loading control. ( c ) Serum-induced cell migration of HMEC transfected with a Cx43-siRNA or a control siRNA (Ctrl) in response to 10% FCS was assessed in a wound assay. Downregulation of Cx43 significantly decreased cell migration, represented as accumulated distance (*** p < 0.001 Ctrl-siRNA vs. Cx43-siRNA, n = 6, 3 independent cell cultures). ( d ) Representative single cell traces of migrated HMEC.

Article Snippet: For immunoprecipitation, the supernatants were incubated over night at 4 °C with a mouse anti-SHP-2 antibody (Santa Cruz) or a rabbit anti-Cx43 antibody (Sigma Aldrich), and were magnetic beads coated with protein A (Cx43) or G (SHP-2) (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Migration, Western Blot, Expressing, Transfection, Control, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: Cx43 Promotes Endothelial Cell Migration and Angiogenesis via the Tyrosine Phosphatase SHP-2

doi: 10.3390/ijms23010294

Figure Lengend Snippet: Downregulation of Cx43 impairs vessel sprouting ex vivo. ( a ) Downregulation of Cx43 in the mouse aortae was achieved by transfection with Cx43 siRNA (500 nM) or control (Ctrl) siRNA, as assessed by Western blot 48 h later. ( b ) Representative images of aortic sprouts in matrigel. Scale bar represents 200 µm. ( c ) The vessel sprouting area (** p < 0.01 Ctrl-siRNA vs. Cx43-siRNA, n = 2 aortae with eight aortic segments each at 3 days; n = 3–4 aortae with 6–8 aortic segments each at 6 days) and ( d ) the number of sprout bifurcations (** p < 0.01 Ctrl-siRNA vs. Cx43-siRNA, n = 2 aortae with 8 aortic segments each at 3 days; *** p < 0.001 Ctrl-siRNA vs. Cx43-siRNA n = 3–4 aortae with 6–8 aortic segments each at 6 days) were significantly reduced upon downregulation of Cx43, as assessed 3 days and 6 days after transfection in a matrigel assay.

Article Snippet: For immunoprecipitation, the supernatants were incubated over night at 4 °C with a mouse anti-SHP-2 antibody (Santa Cruz) or a rabbit anti-Cx43 antibody (Sigma Aldrich), and were magnetic beads coated with protein A (Cx43) or G (SHP-2) (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Ex Vivo, Transfection, Control, Western Blot, Matrigel Assay

Journal: International Journal of Molecular Sciences

Article Title: Cx43 Promotes Endothelial Cell Migration and Angiogenesis via the Tyrosine Phosphatase SHP-2

doi: 10.3390/ijms23010294

Figure Lengend Snippet: Interaction of Cx43 and SHP-2 induces SHP-2 phosphatase activity. ( a ) Upon immunoprecipitation (IP) of SHP-2 from HeLa cells expressing only Cx43, Cx43 was co-immunoprecipitated. As a control, cells transfected with an empty vector (control (CTL)) were used. L: Whole cell lysates. ( b ) SHP-2 was detected upon immunoblotting after immunoprecipitation of Cx43 from HeLa cells expressing only Cx43 in contrast to the control cells (CTL). L: Whole cell lysates. Images originates from the same blot, which was cropped; ( c ) SHP-2 was immunoprecipitated together with Cx43 in HMEC; ( d ) Co-localization of SHP-2 and Cx43 was detected by immunofluorescent staining of HMEC followed by confocal microscopic imaging. Scale bar represents 20 µm; ( e ) SHP-2 phosphatase activity was significantly increased in HeLa cells expressing Cx43 compared to HeLa cells transfected with an empty vector (* p < 0.05, n = 4 independent cell cultures), as assessed by detection of dephosphorylation of a phosphate analogue in SHP-2 immunoprecipitates.

Article Snippet: For immunoprecipitation, the supernatants were incubated over night at 4 °C with a mouse anti-SHP-2 antibody (Santa Cruz) or a rabbit anti-Cx43 antibody (Sigma Aldrich), and were magnetic beads coated with protein A (Cx43) or G (SHP-2) (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Activity Assay, Immunoprecipitation, Expressing, Control, Transfection, Plasmid Preparation, Western Blot, Staining, Imaging, De-Phosphorylation Assay

Journal: International Journal of Molecular Sciences

Article Title: Cx43 Promotes Endothelial Cell Migration and Angiogenesis via the Tyrosine Phosphatase SHP-2

doi: 10.3390/ijms23010294

Figure Lengend Snippet: Inactivation of SHP-2 reduces endothelial cell migration. ( a ) Overexpression of a dominant negative substrate trapping mutant SHP-2 (SHP-2 CS) in HMEC significantly reduced serum (10%) induced endothelial cell migration, as assessed by the accumulated distance in µm in an in vitro wound assay (* p < 0.05, n = 3 independent cell cultures) compared to SHP-2 WT. Additional knock-down of Cx43 (Cx43 siRNA) did not further decrease migration (ns: not significant, n = 3 independent cell cultures). ( b ) Representative single cell traces of migrated HMEC.

Article Snippet: For immunoprecipitation, the supernatants were incubated over night at 4 °C with a mouse anti-SHP-2 antibody (Santa Cruz) or a rabbit anti-Cx43 antibody (Sigma Aldrich), and were magnetic beads coated with protein A (Cx43) or G (SHP-2) (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Migration, Over Expression, Dominant Negative Mutation, Mutagenesis, In Vitro, Knockdown

Journal: International Journal of Molecular Sciences

Article Title: Cx43 Promotes Endothelial Cell Migration and Angiogenesis via the Tyrosine Phosphatase SHP-2

doi: 10.3390/ijms23010294

Figure Lengend Snippet: The Cx43 and SHP-2 interaction is vital for endothelial migration. ( a ) Treatment of HMEC overexpressing a constitutively active mutant SHP-2 (SHP-2 EA) with Cx43 siRNA did not rescue the reduced migratory response caused by Cx43 knock-down (Cx43 siRNA) (ns: not significant, n = 3 independent cell cultures) compared to SHP-2 EA cells treated with control siRNA (Ctrl) (* p < 0.05, n = 3 independent cell cultures), as assessed by the accumulated distance in µm in an in vitro wound assay (* p < 0.05, n = 3 independent cell cultures). ( b ) Representative single cell traces of migrated HMEC.

Article Snippet: For immunoprecipitation, the supernatants were incubated over night at 4 °C with a mouse anti-SHP-2 antibody (Santa Cruz) or a rabbit anti-Cx43 antibody (Sigma Aldrich), and were magnetic beads coated with protein A (Cx43) or G (SHP-2) (Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques: Migration, Mutagenesis, Knockdown, Control, In Vitro

Journal: Journal of Traditional Chinese Medicine

Article Title: Efficacy of Bushenjianpi prescription on autoimmune premature ovarian failure in mice

doi: 10.1016/S0254-6272(17)30321-7

Figure Lengend Snippet: Figure 3 Immunohistochemical analysis of the distinct presence of Cx43 and BMP-15 in the oophorons of each group (× 200) A-F: Cx43; G-L: BMP-15. A, G: control group; B, H: model group; C, I: positive group; D, J: low dose of BSJPP group; E, K: moderate dose of BSJPP group; F, L: high dose of BSJPP group. Positive group treated with premarin (0.03 mg/kg) once daily for 90 d. Con- trol and model group received the same volume of distilled water. Low (8.1 mg/kg), moderate (16.2 mg/kg, the dose used in vivo in human studies), and high (32.4 mg/kg) dose of BSJPP groups treated by gastrogavage once daily for 90 d. Immunohistochemi- cal analysis clearly revealed the presence of Cx43 and BMP-15 in the oophorons of control mice and those treated with BSJPP (D, E) and premarin compared with the model group. BSJPP: Bushenjianpi prescription; Cx43: connexin 43.

Article Snippet: Anti-connexin 43 (Cx43) antibody (product lot No. BA1727) and anti-bone morphogenetic protein 15 (BMP-15) antibody (product lot No. BA2018) were purchased from Boster Biological Engineering (Wuhan, China).

Techniques: Immunohistochemical staining, Control, In Vivo

Journal: Journal of Traditional Chinese Medicine

Article Title: Efficacy of Bushenjianpi prescription on autoimmune premature ovarian failure in mice

doi: 10.1016/S0254-6272(17)30321-7

Figure Lengend Snippet: Figure 4 Expression Cx43 and BMP-15 mRNA in the ovaries of mice in each group. 1: control group; 2: model group; 3: positive group; 4: low dose of BSJPP group; 5: moderate dose of BSJPP group; 6: high dose of BSJPP group. Positive group treated with pre- marin (0.03 mg/kg) once daily for 90 d. Control and model group received the same volume of distilled water. Low (8.1 mg/kg), moderate (16.2 mg/kg, the dose used in vivo in human studies), and high (32.4 mg/kg) dose of BSJPP groups treated by gastrogavage once daily for 90 d. Compared with the model group, significantly upregulated expression of both Cx43 and BMP-15 was detected in the control group and the groups treated with BSJPP (M and L groups). BSJPP: Bushenjianpi prescription; Cx43: connexin 43.

Article Snippet: Anti-connexin 43 (Cx43) antibody (product lot No. BA1727) and anti-bone morphogenetic protein 15 (BMP-15) antibody (product lot No. BA2018) were purchased from Boster Biological Engineering (Wuhan, China).

Techniques: Expressing, Control, In Vivo

Journal: Viruses

Article Title: SARS-CoV-2 Structural Proteins Modulated Blood-Testis Barrier-Related Proteins through Autophagy in the Primary Sertoli Cells.

doi: 10.3390/v15061272

Figure Lengend Snippet: Figure 2. The effects of SARS-CoV-2 SPs on the expression of BTB-related proteins. (A) qPCR analysis of the mRNA expression of BTB-related genes. (A-a–A-f) Sertoli cells were transiently transfected with SARS-CoV-2 SP, then total RNAs were extracted for qPCR to analyze the mRNA expressions of ZO-1, claudin11, occludin, N-cadherin, β-catenin, and CX43. β-actin was the internal control. (B) Immunoblotting analysis of the expressions of BTB-related proteins. (B-a) Sertoli cells were transiently transfected with SARS-CoV-2 SPs, and then total cellular extracts were analyzed by immunoblotting of ZO-1, Claudin11, occludin, N-cadherin, β-Catenin, CX43, and GAPDH as the internal control. (B-b–B-g) The relative levels of the targeted proteins were shown by histograms representing density readings of the gel bands, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of three independent experiments. * p < 0.05, ** p < 0.01, calculated using the Student’s t-test of SARS-CoV-2 SPs transfected cells vs. empty plasmid (GFP-V) transfected cells.

Article Snippet: Rabbit antibodies specific to CX43 (A00599), N-cadherin (BA0673), and β-catenin (BA0426) were purchased from Boster (Wuhan, China).

Techniques: Expressing, Transfection, Control, Western Blot, Plasmid Preparation

Journal: Viruses

Article Title: SARS-CoV-2 Structural Proteins Modulated Blood-Testis Barrier-Related Proteins through Autophagy in the Primary Sertoli Cells.

doi: 10.3390/v15061272

Figure Lengend Snippet: Figure 5. Inhibiting autophagy with 3-MA suppressed SARS-CoV-2 SPs’ effects on BTB-related proteins. (a) Sertoli cells were pretreated with 3-MA (5 mM) for 6 h, then transiently transfected with SARS-CoV-2 SPs. The expression of LC3, ZO-1, claudin11, occludin, N-cadherin, β-catenin, CX43, and GAPDH (internal control) was analyzed by immunoblotting with specific antibodies as described in Materials and Methods. (b–g) The relative levels of the targeted proteins were shown by histograms representing density readings of the gel bands, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of three independent experiments. * p < 0.05, ** p < 0.01, calculated using the Student’s t-test of SARS-CoV-2 SPs transfected cells vs. empty plasmid transfected cells.

Article Snippet: Rabbit antibodies specific to CX43 (A00599), N-cadherin (BA0673), and β-catenin (BA0426) were purchased from Boster (Wuhan, China).

Techniques: Transfection, Expressing, Control, Western Blot, Plasmid Preparation

Journal: Oxidative Medicine and Cellular Longevity

Article Title: DL0410 Alleviates Memory Impairment in D-Galactose-Induced Aging Rats by Suppressing Neuroinflammation via the TLR4/MyD88/NF- κ B Pathway

doi: 10.1155/2021/6521146

Figure Lengend Snippet: DL0410 increased the expression of tight junction proteins and maintained integrity of blood-brain barrier (BBB) in the hippocampus cortex of rats stimulated by D-gal. (a) The representative images and statistical analysis of relative expression for claudin-1, claudin-5, occludin CX43, and ZO-1 in the hippocampus. (b) The representative images and statistical analysis of relative expression for claudin-1, claudin-5, occludin CX43, and ZO-1 in the cortex. Values are mean ± SEM of six independent experiments. # p < 0.05, ## p < 0.01, ### p < 0.001 vs. the control group, ∗ p < 0.05, ∗∗ p < 0.01 vs. the model group.

Article Snippet: Primary antibody against connexin 43 (CX43) (#ER1802-88) was the product of Hangzhou HuaAn Biotechnology Co., Ltd. (Hangzhou, China).

Techniques: Expressing, Control