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Cell Signaling Technology Inc epac1
Epac1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+epac1/us12502382-67-19-20
Average 86 stars, based on 1 article reviews

epac1 - by Bioz Stars, 2026-10

86/100 stars

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Article Title: Nitration of protein phosphatase 2A increases via Epac1/PLCε/CaMKII/HDAC5/iNOS cascade in human endometrial stromal cell decidualization.
Article Snippet: Anti-iNOS was purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, California, USA); anti-PP2Ac, anti-Epac1, antiphospho-CaMKII (Thr286), and anti-CaMKII antibodies were purchased from Cell Signaling Technology, Inc (Danvers, MA, USA), and anti-phospho-HDAC5 (Ser498) and anti-HDAC5 antibodies were purchased from Abcam (Cambridge, MA, USA).

Incubation:

Article Title: Upregulation of Epac1 Promotes Pericyte Loss by Inducing Mitochondrial Fission, Reactive Oxygen Species Production, and Apoptosis
Article Snippet: .. Subsequently, the vasculatures were incubated with anti-Epac1 (Cat. 4155S; Cell Signaling Technology, Danvers, MA, USA), anti-NG2 (Cat. AB5320; MilliporeSigma, Burlington, MA, USA), and isolectin B4 (Cat. I21412; Invitrogen, Waltham, MA, USA) at 4°C overnight. ..

Article Title: Epac2 Deficiency Compromises Adaptation to Dietary Acidification by Decreasing H + Transport in the Renal Nephron
Article Snippet: .. Nitrocellulose membranes were incubated with primary antibodies overnight at 4°C: anti-Epac1 (mouse mAb, 1:1000 Cell Signaling Technology, USA; Cat. #4155), anti-Epac2 (rabbit mAb, 1:250 Cell Signaling Technology, USA; Cat. #43239), anti-NHE-3 (rabbit mAb, 1:1000 Alomone Labs, Israel; Cat. #ANX-033), anti-NBCe1 (rabbit mAb, 1:1000 Alomone Labs, Israel; Cat. #ANT-07), anti-AE1 (rabbit mAb, 1:1000 Invitrogen, USA; Cat. # PA5-80030), or anti-pendrin (rabbit mAb, 1:1000 Invitrogen, USA; Cat. # PA5-42060). .. Following washout (3 times for 10 min in TBS-Tween), the membrane was incubated with peroxidase-conjugated goat anti-rabbit or anti-mouse (1:10 000, Jackson ImmunoResearch Laboratories, USA) secondary antibodies for 1 h at room temperature.

Article Title: Epac2 deficiency compromises adaptation to dietary acidification by decreasing H+ transport in the renal nephron.
Article Snippet: .. Nitrocellulose membranes were incubated with primary antibodies overnight at 4°C: anti-Epac1 (mouse mAb, 1:1000 Cell Signaling Technology, USA; Cat. # 4155), anti-Epac2 (rabbit mAb, 1:250 Cell Signaling Technology, USA; Cat. # 43239), anti-NHE3 (rabbit mAb, 1:1000 Alomone Labs, Israel; Cat. # ANX-033), anti-NBCe1 (rabbit mAb, 1:1000 Alomone Labs, Israel; Cat. # ANT-07), anti-AE1 (rabbit mAb, 1:1000 Invitrogen, USA; Cat. # PA580030), or anti-pendrin (rabbit mAb, 1:1000 Invitrogen, USA; Cat. # PA5-42060). .. Following washout (3 times for 10 min in TBS-Tween), the membrane was incubated with peroxidase-conjugated goat anti-rabbit or anti-mouse (1:10000, Jackson ImmunoResearch Laboratories, USA) secondary antibodies for 1 hour at room temperature.

Article Title: Upregulation of Epac1 Promotes Pericyte Loss by Inducing Mitochondrial Fission, Reactive Oxygen Species Production, and Apoptosis.
Article Snippet: .. Subsequently, the vasculatures were incubated with anti-Epac1 (Cat. 4155S; Cell Signaling Technology, Danvers, MA, USA), anti-NG2 (Cat. AB5320; MilliporeSigma, Burlington, MA, USA), and isolectin B4 (Cat. I21412; Invitrogen, Waltham, MA, USA) at 4°C overnight. ..

Article Title: Effect of zearalenone on the jejunum of weaned gilts through the Epac1/Rap1/JNK pathway.
Article Snippet: Zearalenone (ZEN) is a nonsteroidal estrogenic mycotoxin produced by Fusarium strains that is harmful to the intestinal health of animals and is widely present in contaminated crops.. The objective of this study was to investigate the potential therapeutic target of ZEN-induced jejunal damage in weaned gilts.. Sixteen weaned gilts either received a basal diet or a basal diet supplemented with 3.0 mg/kg ZEN in a 32-d experiment.

Western Blot:

Article Title: Protein SUMOylation promotes cAMP-independent EPAC1 activation
Article Snippet: .. The signals of EPAC1 PTM, α-Tubulin, and Lamin B were detected by Western Blot using anti-EPAC1 (Cell Signaling Technology), anti-α-Tubulin (Cell Signaling Technology), and anti-Lamin B (Santa Cruz Biotechnology Inc.) antibodies. ..

Cell-Signaling:

Polyacrylamide Gel Electrophoresis:

Article Title: Urocortin 2 promotes hypertrophy and enhances skeletal muscle function through cAMP and insulin/IGF-1 signaling pathways
Article Snippet: .. Lysates were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotted with anti-phospho-Ser 133 CREB (1:1000, Cell Signaling #9198), anti-PGC1-α (1:500, Cell Signaling #2178), anti-β-actin (1:2000, Santa Cruz sc - 81,178), anti-Epac1 (1:500, Cell Signaling #4155), anti-puromycin (1:500, Kerafast EQ 0001), anti-phospho-Ser 2448 mTOR (1:500, Cell Signaling #2971), anti-phospho-Ser 235/236 S6 (1:1000, Cell Signaling #2211), anti-phospho-Thr 37/46 4 E-BP1 (1:1000, Cell Signaling #2855), anti-phospho-Ser 209 eIF4E (1:1000, Cell Signaling #9741), anti-mTOR (1:500, Cell Signaling #2972), anti-S6 (1:500, Cell Signaling #2217), anti-4E-BP1 (1:1000, Cell Signaling #9452), anti-eIF4E (1:1000, Cell Signaling #9742), anti-LC3 (1:1000, Cell Signaling #2775), anti-p62 (1:1000, Cell Signaling #5114), anti-phospho-Ser 256 FoxO1 (1:500, Cell Signaling #9461), anti-phospho-Thr 24 FoxO1 and anti-phospho-Thr 32 FoxO3a (1:500, Cell Signaling #9464), anti-FoxO1 (1:500, Cell Signaling #9454), anti-phospho-Ser 473 Akt (1:1000, Cell Signaling #9271), anti-phospho-Thr 308 Akt (1:500, Cell Signaling #9275), anti-phospho-Thr 202 /Tyr 204 ERK1 and anti-phospho-Thr 185 /Tyr 187 ERK2 (1:1000, Cell Signaling #9101), anti-phospho-Thr 180 /Tyr 182 p38 MAPK (1:1000, Cell Signaling #9211), anti-Akt (1:500, Cell Signaling #9272), anti-ERK1/2 (1:1000, Cell Signaling #9102), anti-slow myosin (1:1000, Sigma–Aldrich M8421), anti-fast myosin (1:2000, Sigma–Aldrich M4276), anti-GFP (1:1000, Santa Cruz sc - 8334), anti-phospho-Tyr 1135 / 1136 IGF-I receptor β and anti-phospho-Tyr 1150 / 1151 Insulin Receptor β (1:500, Cell Signaling #3024). .. Primary antibodies were detected using peroxidase-conjugated secondary antibodies (1:5000 for β-actin, slow- and fast-myosin and 1:1000 for the other primary antibodies) and visualized using enhanced chemiluminescence (ECL) reagents on ChemiDoc XRS + System (Bio-Rad).

Image Search Results

Epac1 is downregulated in orbital tissues of patients with TAO . ( A – E ) The collected orbital tissue samples from patients with TAO and healthy donors were assessed for histopathological features using H&E and Masson stainings ( A , B ); Epac1 level and distribution using IHC staining ( C ); Epac1 mRNA expression using qRT-PCR ( D ); and Epac1 and vimentin protein levels using immunoblotting ( E ) ( n = 6 or 5). The Student's t -test was conducted to assess comparisons between the two groups, ** P < 0.01 versus the healthy group. TAO, thyroid-associated orbitopathy; H&E, hematoxylin and eosin staining; IHC, immunohistochemical staining; OAT, orbital adipose tissues.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Epac1 Inhibits Orbital Fibroblast Activation to Ameliorate Thyroid-Associated Orbitopathy-Like Features Through the JAK/STAT Signaling Pathway

doi: 10.1167/iovs.66.9.68

Figure Lengend Snippet: Epac1 is downregulated in orbital tissues of patients with TAO . ( A – E ) The collected orbital tissue samples from patients with TAO and healthy donors were assessed for histopathological features using H&E and Masson stainings ( A , B ); Epac1 level and distribution using IHC staining ( C ); Epac1 mRNA expression using qRT-PCR ( D ); and Epac1 and vimentin protein levels using immunoblotting ( E ) ( n = 6 or 5). The Student's t -test was conducted to assess comparisons between the two groups, ** P < 0.01 versus the healthy group. TAO, thyroid-associated orbitopathy; H&E, hematoxylin and eosin staining; IHC, immunohistochemical staining; OAT, orbital adipose tissues.

Article Snippet: The used primary antibodies were α-SMA (55135-1-AP; Proteintech), Collagen I (14695-1-AP; Proteintech), Vimentin (10366-1-AP; Proteintech), Epac1 (DF6922; Affinity), fibronectin (15613-1-AP; Proteintech), collagen III (22734-1-AP; Proteintech), phosphorylated STAT3 (CSB-RA022812A727phHU; Cusabio, Wuhan, China), STAT3 (10253-2-AP; Proteintech), FABP4 (12802-1-AP; Protientech), and β-actin (CSB- MA000187 ; Cusabio).

Techniques: Immunohistochemistry, Expressing, Quantitative RT-PCR, Western Blot, Staining, Immunohistochemical staining

Epac1 is downregulated in OAT and OMT in a TAO mouse model. Mouse OAT and OMT samples were evaluated for histopathological features using H&E and Masson stainings ( A , B ); Epac1 level and distribution using IHC staining ( C ); Epac1 mRNA expression using qRT-PCR ( D ); and Epac1 and vimentin protein levels using immunoblotting ( E ) ( n = 5). The Student's t -test was conducted to assess comparisons between the two groups, ** P < 0.01 versus the Ad-NC group. H&E, hematoxylin and eosin staining; IHC, immunohistochemical staining; OAT, orbital adipose tissues; OMT, orbital muscle tissues; Ad, adenovirus; NC, negative control.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Epac1 Inhibits Orbital Fibroblast Activation to Ameliorate Thyroid-Associated Orbitopathy-Like Features Through the JAK/STAT Signaling Pathway

doi: 10.1167/iovs.66.9.68

Figure Lengend Snippet: Epac1 is downregulated in OAT and OMT in a TAO mouse model. Mouse OAT and OMT samples were evaluated for histopathological features using H&E and Masson stainings ( A , B ); Epac1 level and distribution using IHC staining ( C ); Epac1 mRNA expression using qRT-PCR ( D ); and Epac1 and vimentin protein levels using immunoblotting ( E ) ( n = 5). The Student's t -test was conducted to assess comparisons between the two groups, ** P < 0.01 versus the Ad-NC group. H&E, hematoxylin and eosin staining; IHC, immunohistochemical staining; OAT, orbital adipose tissues; OMT, orbital muscle tissues; Ad, adenovirus; NC, negative control.

Techniques: Immunohistochemistry, Expressing, Quantitative RT-PCR, Western Blot, Staining, Immunohistochemical staining, Negative Control

Epac1 overexpression attenuates the effects of TGFβ1 on normal and TAO OFs. ( A – G ) Healthy or TAO OFs were transfected with Epac1 -overexpressing plasmid ( Epac1 ), treated with TGFβ1 (10 ng/mL), and examined for cell viability using CCK-8 ( A ); cell migration using scratch wound healing and Transwell assays ( B , C ); relative α-SMA and fibronectin expressions using IF staining ( D ); collagen I content in the cell culture supernatant using ELISA ( E ); the mRNA expression of Epac1 , α-SMA, vimentin, fibronectin, collagen I, and collagen III using qRT-PCR ( F ); and the protein level of Epac1, α-SMA, vimentin, fibronectin, collagen I, and collagen III using immunoblotting ( G ) ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. ** P < 0.01 versus healthy OFs; # P < 0.05, ## P < 0.01 versus TAO OFs; && P < 0.01 versus healthy OFs + TGFβ1 + NC; $ P < 0.05, $$ P < 0.01 versus TAO OFs + TGFβ1 + NC. TAO, thyroid-associated orbitopathy; sh, short-hairpin; NC, negative control; IF staining, immunofluorescent staining.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Epac1 Inhibits Orbital Fibroblast Activation to Ameliorate Thyroid-Associated Orbitopathy-Like Features Through the JAK/STAT Signaling Pathway

doi: 10.1167/iovs.66.9.68

Figure Lengend Snippet: Epac1 overexpression attenuates the effects of TGFβ1 on normal and TAO OFs. ( A – G ) Healthy or TAO OFs were transfected with Epac1 -overexpressing plasmid ( Epac1 ), treated with TGFβ1 (10 ng/mL), and examined for cell viability using CCK-8 ( A ); cell migration using scratch wound healing and Transwell assays ( B , C ); relative α-SMA and fibronectin expressions using IF staining ( D ); collagen I content in the cell culture supernatant using ELISA ( E ); the mRNA expression of Epac1 , α-SMA, vimentin, fibronectin, collagen I, and collagen III using qRT-PCR ( F ); and the protein level of Epac1, α-SMA, vimentin, fibronectin, collagen I, and collagen III using immunoblotting ( G ) ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. ** P < 0.01 versus healthy OFs; # P < 0.05, ## P < 0.01 versus TAO OFs; && P < 0.01 versus healthy OFs + TGFβ1 + NC; $ P < 0.05, $$ P < 0.01 versus TAO OFs + TGFβ1 + NC. TAO, thyroid-associated orbitopathy; sh, short-hairpin; NC, negative control; IF staining, immunofluorescent staining.

Techniques: Over Expression, Transfection, Plasmid Preparation, CCK-8 Assay, Migration, Staining, Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Western Blot, Negative Control

Epac1 knockdown amplifies the effects of TGFβ1 on normal and TAO OFs. ( A , B ) Healthy or TAO OFs were transfected with plasmids containing short-hairpin RNA targeting Epac1 (sh- Epac1 #1/#2), treated with TGFβ1 (10 ng/mL), and examined for Epac1 protein levels using immunoblotting; and ( B ) cell viability using CCK-8; sh- Epac1 #1 was selected for the following experiments. ( C – H ) Healthy or TAO OFs were transfected with sh- Epac1 #1, treated with TGFβ1 (10 ng/mL), and examined for cell migration using scratch wound healing and Transwell assays ( C , D ); relative expression of α-SMA and fibronectin using IF staining ( E ); the content of collagen I in cell culture supernatant using ELISA (F); the mRNA expression of Epac1 , α-SMA, vimentin, fibronectin, collagen I, and collagen III using qRT-PCR ( G ); and the protein level of Epac1, α-SMA, vimentin, fibronectin, collagen I, and collagen III using immunoblotting ( H ) ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. * P < 0.05, ** P < 0.01 versus healthy OFs + TGFβ1 + sh-NC; # P < 0.05, ## P < 0.01 versus TAO OFs + TGFβ1 + sh-NC. TAO, thyroid-associated orbitopathy; sh, short hairpin RNA; NC, negative control; IF staining, immunofluorescent staining.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Epac1 Inhibits Orbital Fibroblast Activation to Ameliorate Thyroid-Associated Orbitopathy-Like Features Through the JAK/STAT Signaling Pathway

doi: 10.1167/iovs.66.9.68

Figure Lengend Snippet: Epac1 knockdown amplifies the effects of TGFβ1 on normal and TAO OFs. ( A , B ) Healthy or TAO OFs were transfected with plasmids containing short-hairpin RNA targeting Epac1 (sh- Epac1 #1/#2), treated with TGFβ1 (10 ng/mL), and examined for Epac1 protein levels using immunoblotting; and ( B ) cell viability using CCK-8; sh- Epac1 #1 was selected for the following experiments. ( C – H ) Healthy or TAO OFs were transfected with sh- Epac1 #1, treated with TGFβ1 (10 ng/mL), and examined for cell migration using scratch wound healing and Transwell assays ( C , D ); relative expression of α-SMA and fibronectin using IF staining ( E ); the content of collagen I in cell culture supernatant using ELISA (F); the mRNA expression of Epac1 , α-SMA, vimentin, fibronectin, collagen I, and collagen III using qRT-PCR ( G ); and the protein level of Epac1, α-SMA, vimentin, fibronectin, collagen I, and collagen III using immunoblotting ( H ) ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. * P < 0.05, ** P < 0.01 versus healthy OFs + TGFβ1 + sh-NC; # P < 0.05, ## P < 0.01 versus TAO OFs + TGFβ1 + sh-NC. TAO, thyroid-associated orbitopathy; sh, short hairpin RNA; NC, negative control; IF staining, immunofluorescent staining.

Techniques: Knockdown, Transfection, shRNA, Western Blot, CCK-8 Assay, Migration, Expressing, Staining, Cell Culture, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Negative Control

Epac1 overexpression improves TAO-like features in the mouse model. ( A ) A schematic diagram of the TAO model establishment and Epac1 overexpression administration; ( B ) at the end of the modeling, the appearance of mouse eyes was observed; ( C , D ) the histopathological alterations in mouse OAT and OMT samples were evaluated using H&E and Masson stainings. TAO, thyroid-associated orbitopathy; OAT, orbital adipose tissues; OMT, orbital muscle tissues; AAV, adeno-associated virus; NC, negative control; L, left; R, right; H&E, hematoxylin and eosin staining.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Epac1 Inhibits Orbital Fibroblast Activation to Ameliorate Thyroid-Associated Orbitopathy-Like Features Through the JAK/STAT Signaling Pathway

doi: 10.1167/iovs.66.9.68

Figure Lengend Snippet: Epac1 overexpression improves TAO-like features in the mouse model. ( A ) A schematic diagram of the TAO model establishment and Epac1 overexpression administration; ( B ) at the end of the modeling, the appearance of mouse eyes was observed; ( C , D ) the histopathological alterations in mouse OAT and OMT samples were evaluated using H&E and Masson stainings. TAO, thyroid-associated orbitopathy; OAT, orbital adipose tissues; OMT, orbital muscle tissues; AAV, adeno-associated virus; NC, negative control; L, left; R, right; H&E, hematoxylin and eosin staining.

Techniques: Over Expression, Virus, Negative Control, Staining

Epac1 overexpression decreases TAO-associated markers in vivo. ( A , B ) The protein level of Epac1, α-SMA, CD40, and collagen I in mouse OAT was examined using IHC staining; ( C , D ) the protein level of Epac1, α-SMA, CD40, and collagen I in mouse OMT was examined using IHC staining ( n = 5). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. * P < 0.05, ** P < 0.01 versus healthy; ## P < 0.01 versus TAO + AAV-NC. TAO, thyroid-associated orbitopathy; OAT, orbital adipose tissues; OMT, orbital muscle tissues; AAV, adeno-associated virus; NC, negative control; IHC staining, immunohistochemical staining.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Epac1 Inhibits Orbital Fibroblast Activation to Ameliorate Thyroid-Associated Orbitopathy-Like Features Through the JAK/STAT Signaling Pathway

doi: 10.1167/iovs.66.9.68

Figure Lengend Snippet: Epac1 overexpression decreases TAO-associated markers in vivo. ( A , B ) The protein level of Epac1, α-SMA, CD40, and collagen I in mouse OAT was examined using IHC staining; ( C , D ) the protein level of Epac1, α-SMA, CD40, and collagen I in mouse OMT was examined using IHC staining ( n = 5). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. * P < 0.05, ** P < 0.01 versus healthy; ## P < 0.01 versus TAO + AAV-NC. TAO, thyroid-associated orbitopathy; OAT, orbital adipose tissues; OMT, orbital muscle tissues; AAV, adeno-associated virus; NC, negative control; IHC staining, immunohistochemical staining.

Techniques: Over Expression, In Vivo, Immunohistochemistry, Virus, Negative Control, Immunohistochemical staining, Staining

Epac1 affects the STAT3 signaling. ( A ) A schematic diagram of Epac1’s role in the JAK/STAT3 signaling pathway during the OF activation process. ( B ) TAO OFs transfected with sh- Epac1 or Epacl overexpression vector, treated with TGFβ1, and determined for the protein level of p-STAT3 and STAT3 using immunoblotting ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. ** P < 0.01 versus NC-transfected OFs; ## P < 0.01 versus sh-NC-transfected OFs. ( C ) The protein level of p-STAT3 and STAT3 in TAO mouse OAT and OMT was determined using immunoblotting ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. ** P < 0.01 versus healthy mice; # P < 0.05, ## P < 0.01 versus TAO + AAV-NC mice. TAO, thyroid-associated orbitopathy; OAT, orbital adipose tissues; OMT, orbital muscle tissues; AAV, adeno-associated virus; sh, short-hairpin RNA; NC, negative control.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Epac1 Inhibits Orbital Fibroblast Activation to Ameliorate Thyroid-Associated Orbitopathy-Like Features Through the JAK/STAT Signaling Pathway

doi: 10.1167/iovs.66.9.68

Figure Lengend Snippet: Epac1 affects the STAT3 signaling. ( A ) A schematic diagram of Epac1’s role in the JAK/STAT3 signaling pathway during the OF activation process. ( B ) TAO OFs transfected with sh- Epac1 or Epacl overexpression vector, treated with TGFβ1, and determined for the protein level of p-STAT3 and STAT3 using immunoblotting ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. ** P < 0.01 versus NC-transfected OFs; ## P < 0.01 versus sh-NC-transfected OFs. ( C ) The protein level of p-STAT3 and STAT3 in TAO mouse OAT and OMT was determined using immunoblotting ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. ** P < 0.01 versus healthy mice; # P < 0.05, ## P < 0.01 versus TAO + AAV-NC mice. TAO, thyroid-associated orbitopathy; OAT, orbital adipose tissues; OMT, orbital muscle tissues; AAV, adeno-associated virus; sh, short-hairpin RNA; NC, negative control.

Techniques: Activation Assay, Transfection, Over Expression, Plasmid Preparation, Western Blot, Virus, shRNA, Negative Control

STAT3 mediates the effects of Epac1 on TGFβ1-treated TAO OFs. ( A – G ) Under TGFβ1 treatment, TAO OFs were transfected with sh- Epac1 with or without the JAK-STAT pathway inhibitor Stattic (2 g/mL for 24 hours) and examined for cell viability using CCK-8 ( A ); cell migration using scratch wound healing and Transwell assays ( B , C ); α-SMA and fibronectin expressions using IF staining ( D ); the content of collagen I in cell supernatant using ELISA ( E ); the mRNA expression of Epac1 , α-SMA, vimentin, fibronectin, collagen I, and collagen III using qRT-PCR ( F ); the protein level of α-SMA, vimentin, fibronectin, collagen I, and collagen III using immunoblotting ( G ) ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. ** P < 0.01 versus TGFβ1 + sh-NC; ## P < 0.01 versus TGFβ1 + sh- Epac1 + Stattic. sh, short-hairpin; NC, negative control; IF staining, immunofluorescent staining.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Epac1 Inhibits Orbital Fibroblast Activation to Ameliorate Thyroid-Associated Orbitopathy-Like Features Through the JAK/STAT Signaling Pathway

doi: 10.1167/iovs.66.9.68

Figure Lengend Snippet: STAT3 mediates the effects of Epac1 on TGFβ1-treated TAO OFs. ( A – G ) Under TGFβ1 treatment, TAO OFs were transfected with sh- Epac1 with or without the JAK-STAT pathway inhibitor Stattic (2 g/mL for 24 hours) and examined for cell viability using CCK-8 ( A ); cell migration using scratch wound healing and Transwell assays ( B , C ); α-SMA and fibronectin expressions using IF staining ( D ); the content of collagen I in cell supernatant using ELISA ( E ); the mRNA expression of Epac1 , α-SMA, vimentin, fibronectin, collagen I, and collagen III using qRT-PCR ( F ); the protein level of α-SMA, vimentin, fibronectin, collagen I, and collagen III using immunoblotting ( G ) ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. ** P < 0.01 versus TGFβ1 + sh-NC; ## P < 0.01 versus TGFβ1 + sh- Epac1 + Stattic. sh, short-hairpin; NC, negative control; IF staining, immunofluorescent staining.

Techniques: Transfection, CCK-8 Assay, Migration, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Western Blot, Negative Control

Effects of EphrinB2 overexpression on Epac1-Rap1 Pathway in diabetic nephropathy mouse. A Immunofluorescence staining of EphrinB2 (red) and Epac1 (green) in kidney tissues from different experimental groups. DAPI (blue) was used for nuclear staining. Merged images show colocalization. Scale bar = 50 μm. B , C Quantification of EphrinB2 fluorescence intensity ( B ) and Epac1 fluorescence intensity ( C ) in different groups. ( D ) Immunohistochemical staining of Epac1, Rap1, and PGC-1α in kidney tissues from control (CTL) and STZ-induced diabetic mice, with or without EphrinB2 overexpression. Scale bar = 50 μm. E–G Quantification of Epac1-positive area ( E ), Rap1-positive area ( F ), and PGC-1α-positive area ( G ) in different groups. ( H ) Western blot analysis of Epac1 and Rap1 protein expression in kidney tissues from wild-type (WT), STZ 12-week (STZ12W), and STZ 16-week (STZ16W) mice. β-actin was used as a loading control. I Densitometric quantification of Epac1 and Rap1 protein levels, normalized to β-actin, in WT, STZ12W, and STZ16W groups. All results were presented as the mean ± SD, n = 3/group. * p < 0.05, ** p < 0.01

Journal: Journal of Translational Medicine

Article Title: EphrinB2 alleviates tubulointerstitial fibrosis in diabetic kidney disease

doi: 10.1186/s12967-025-06852-1

Figure Lengend Snippet: Effects of EphrinB2 overexpression on Epac1-Rap1 Pathway in diabetic nephropathy mouse. A Immunofluorescence staining of EphrinB2 (red) and Epac1 (green) in kidney tissues from different experimental groups. DAPI (blue) was used for nuclear staining. Merged images show colocalization. Scale bar = 50 μm. B , C Quantification of EphrinB2 fluorescence intensity ( B ) and Epac1 fluorescence intensity ( C ) in different groups. ( D ) Immunohistochemical staining of Epac1, Rap1, and PGC-1α in kidney tissues from control (CTL) and STZ-induced diabetic mice, with or without EphrinB2 overexpression. Scale bar = 50 μm. E–G Quantification of Epac1-positive area ( E ), Rap1-positive area ( F ), and PGC-1α-positive area ( G ) in different groups. ( H ) Western blot analysis of Epac1 and Rap1 protein expression in kidney tissues from wild-type (WT), STZ 12-week (STZ12W), and STZ 16-week (STZ16W) mice. β-actin was used as a loading control. I Densitometric quantification of Epac1 and Rap1 protein levels, normalized to β-actin, in WT, STZ12W, and STZ16W groups. All results were presented as the mean ± SD, n = 3/group. * p < 0.05, ** p < 0.01

Article Snippet: Epac1 , 1:1000 , Santa Cruz , SC-28366.

Techniques: Over Expression, Immunofluorescence, Staining, Fluorescence, Immunohistochemical staining, Control, Western Blot, Expressing

EphrinB2 Overexpression Restores Mitochondrial Dynamics and Reduces Fibrosis via the Epac1-Rap1 Pathway. A Western blot analysis of proteins involved in mitochondrial dynamics and function, including p-Drp1, Drp1, Opa1, Mfn1, Mfn2, PGC-1α, Rap1, and Epac1 in control and STZ-treated mice with or without EphrinB2 overexpression (EphrinB2-OE). β-actin serves as a loading control. B Quantification of protein expression for EPAC, Rap1, and PGC-1α showing relative changes across different experimental groups. C Quantification of mitochondrial fusion proteins Mfn1, Mfn2, and Opa1 expression levels. D Ratio of phosphorylated Drp1 (p-Drp1) to total Drp1 indicating changes in mitochondrial fission upon different treatments. E Western blot analysis for the same set of proteins in cultured cells under low glucose (LG) and high glucose (HG) conditions, with or without EphrinB2-OE. F Quantification of EphrinB2 and PGC-1α protein levels under different glucose conditions. G Bar graphs showing relative protein levels of EPAC and Rap1 in cultured cells under high glucose conditions. H Expression levels of mitochondrial fusion proteins Mfn1, Mfn2, and Opa1 under different experimental setups. I Analysis of p-Drp1 levels in cells exposed to high glucose with or without EphrinB2-OE. J Western blots for fibrosis markers including Fibronectin and α-SMA, along with mitochondrial dynamics proteins under various conditions. K – L Quantification of EPAC1, Rap1, and PGC-1α levels under the influence of high glucose and different treatment modifiers. ( M ) . Expression levels of mitochondrial fusion proteins under low and high glucose conditions with additional treatment conditions. N Quantitative analysis of α-SMA levels, showing the effect of EphrinB2-OE and its modifiers on fibrotic response in high glucose conditions. O , P Statistical analysis of the ratio of p-Drp1 to Drp1 and the expression levels of fibrosis markers α-SMA and Fibronectin, indicating the regulatory impact of EphrinB2 on mitochondrial dynamics and cellular fibrosis under diabetic conditions. All results were presented as the mean ± SD, n = 3/group. * p < 0.05, ** p < 0.01

Journal: Journal of Translational Medicine

Article Title: EphrinB2 alleviates tubulointerstitial fibrosis in diabetic kidney disease

doi: 10.1186/s12967-025-06852-1

Figure Lengend Snippet: EphrinB2 Overexpression Restores Mitochondrial Dynamics and Reduces Fibrosis via the Epac1-Rap1 Pathway. A Western blot analysis of proteins involved in mitochondrial dynamics and function, including p-Drp1, Drp1, Opa1, Mfn1, Mfn2, PGC-1α, Rap1, and Epac1 in control and STZ-treated mice with or without EphrinB2 overexpression (EphrinB2-OE). β-actin serves as a loading control. B Quantification of protein expression for EPAC, Rap1, and PGC-1α showing relative changes across different experimental groups. C Quantification of mitochondrial fusion proteins Mfn1, Mfn2, and Opa1 expression levels. D Ratio of phosphorylated Drp1 (p-Drp1) to total Drp1 indicating changes in mitochondrial fission upon different treatments. E Western blot analysis for the same set of proteins in cultured cells under low glucose (LG) and high glucose (HG) conditions, with or without EphrinB2-OE. F Quantification of EphrinB2 and PGC-1α protein levels under different glucose conditions. G Bar graphs showing relative protein levels of EPAC and Rap1 in cultured cells under high glucose conditions. H Expression levels of mitochondrial fusion proteins Mfn1, Mfn2, and Opa1 under different experimental setups. I Analysis of p-Drp1 levels in cells exposed to high glucose with or without EphrinB2-OE. J Western blots for fibrosis markers including Fibronectin and α-SMA, along with mitochondrial dynamics proteins under various conditions. K – L Quantification of EPAC1, Rap1, and PGC-1α levels under the influence of high glucose and different treatment modifiers. ( M ) . Expression levels of mitochondrial fusion proteins under low and high glucose conditions with additional treatment conditions. N Quantitative analysis of α-SMA levels, showing the effect of EphrinB2-OE and its modifiers on fibrotic response in high glucose conditions. O , P Statistical analysis of the ratio of p-Drp1 to Drp1 and the expression levels of fibrosis markers α-SMA and Fibronectin, indicating the regulatory impact of EphrinB2 on mitochondrial dynamics and cellular fibrosis under diabetic conditions. All results were presented as the mean ± SD, n = 3/group. * p < 0.05, ** p < 0.01

Article Snippet: Epac1 , 1:1000 , Santa Cruz , SC-28366.

Techniques: Over Expression, Western Blot, Control, Expressing, Cell Culture

EphrinB2 Overexpression Stabilizes Epac1 Protein Under High-Glucose Conditions. A Immunofluorescence staining for EphrinB2 (red) and Epac1 (green) with nuclei stained using DAPI (blue) in cells under low glucose (LG) and high glucose (HG) conditions, with or without EphrinB2 overexpression (EphrinB2-OE). Scale bar, 50 µm. B Quantification of fluorescence intensity for EphrinB2 and Epac1 under the described conditions. C Western blot analysis of Epac1 protein stability with cycloheximide (CHX) treatment over time (0, 4, 8, 12 h) in cells transfected with either vector control or EphrinB2-OE. D Co-immunoprecipitation (co-IP) showing the interaction between EphrinB2 and Epac1 in LG and HG conditions, with or without EphrinB2-OE. E Graph showing the relative expression levels of Epac1 protein over time with CHX treatment in vector control and EphrinB2-OE conditions, demonstrating changes in protein stability. All results were presented as the mean ± SD, n = 3/group. * p < 0.05, ** p < 0.01

Journal: Journal of Translational Medicine

Article Title: EphrinB2 alleviates tubulointerstitial fibrosis in diabetic kidney disease

doi: 10.1186/s12967-025-06852-1

Figure Lengend Snippet: EphrinB2 Overexpression Stabilizes Epac1 Protein Under High-Glucose Conditions. A Immunofluorescence staining for EphrinB2 (red) and Epac1 (green) with nuclei stained using DAPI (blue) in cells under low glucose (LG) and high glucose (HG) conditions, with or without EphrinB2 overexpression (EphrinB2-OE). Scale bar, 50 µm. B Quantification of fluorescence intensity for EphrinB2 and Epac1 under the described conditions. C Western blot analysis of Epac1 protein stability with cycloheximide (CHX) treatment over time (0, 4, 8, 12 h) in cells transfected with either vector control or EphrinB2-OE. D Co-immunoprecipitation (co-IP) showing the interaction between EphrinB2 and Epac1 in LG and HG conditions, with or without EphrinB2-OE. E Graph showing the relative expression levels of Epac1 protein over time with CHX treatment in vector control and EphrinB2-OE conditions, demonstrating changes in protein stability. All results were presented as the mean ± SD, n = 3/group. * p < 0.05, ** p < 0.01

Article Snippet: Epac1 , 1:1000 , Santa Cruz , SC-28366.

Techniques: Over Expression, Immunofluorescence, Staining, Fluorescence, Western Blot, Transfection, Plasmid Preparation, Control, Immunoprecipitation, Co-Immunoprecipitation Assay, Expressing

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