atf3 protein  (TargetMol)

Journal: Biology Direct

Article Title: Platelet-activating factor induces ferroptosis by binding to ATF3 and inhibiting the SLC7A11/GPX4 axis to suppress the progression of endometrial carcinoma

doi: 10.1186/s13062-025-00713-z

Figure Lengend Snippet: PAF induces ferroptosis via the ATF3/GPX4 and ATF3/SLC7A11 axes. (A) Heat map representing the significantly regulated transcription factors detected via RNA-seq analysis of AN3CA and HEC1B cells. (B , C) The mRNA expression levels of ATF3 and DDIT3 were in AN3CA and HEC1B cells treated with PAF (60 µM) for 48 h. (D , E) Protein expression of ATF3 in AN3CA and HEC1B cells treated with PAF (0, 20, 40, and 60 µM) for 48 h (D), and the quantifiable data are shown (E). (F) Confocal microscopy of AN3CA and HEC1B cells in the presence of PAF (60 µM). Cells were stained for ATF3 (green) and phalloidin (red). DAPI was used to visualize the nucleus (blue). Scale bar, 50 μm (left) or 100 μm (right). (G) The potential binding sites between ATF3 and GPX4/SLC7A11 were predicted by JASPAR. (H) Schematic diagram of ATF3 binding site-mutated SLC7A11 reporter vector (pro-SLC7A11-mut) and GPX4 reporter vector (pro-GPX4-mut). (I , J) Dual luciferase assay of the luciferase activity of AN3CA (I) and HEC1B (J) cells. (K) Immunoblotting analysis of the expression of GPX4 and SLC7A11 following ATF3 overexpression in AN3CA and HEC1B cells. (L) Immunoblotting analysis of the protein expression of SLC7A11, GPX4, and ATF3 after ATF3 silencing with siRNA coupled with PAF treatment in AN3CA cells. * p < 0.05, ** p < 0.01 and *** p < 0.001, ns . not significant

Article Snippet: SPR assays were performed by TopScience Co., Ltd. (Shanghai, China) to detect the binding between the recombinant ATF3 protein (TMPH-01164, TargetMol) and C16-PAF (74389-68-7, TargetMol), with the ATF3 protein and anti-ATF3 antibody combination used as the positive control.

Techniques: RNA Sequencing, Expressing, Confocal Microscopy, Staining, Binding Assay, Plasmid Preparation, Luciferase, Activity Assay, Western Blot, Over Expression

Journal: Biology Direct

Article Title: Platelet-activating factor induces ferroptosis by binding to ATF3 and inhibiting the SLC7A11/GPX4 axis to suppress the progression of endometrial carcinoma

doi: 10.1186/s13062-025-00713-z

Figure Lengend Snippet: PAF binds to ATF3 and reduces its ubiquitination. (A , B) Molecular docking indicates the binding details between PAF and ATF3. The surface representation of the protein residues ( A ) and 2D representation of the binding interaction of PAF and ATF3 ( B ) are depicted. (C , D) Surface plasmon resonance of the affinity of anti-ATF3 antibody ( C ) and PAF ( D ) for ATF3 protein. K D , dissociation constant. (E) WB analysis shows that PAF stabilized ATF3 across different temperature gradients in the CETSA in 293T cells. (F) WB analysis indicates that PAF promoted the resistance of ATF3 to pronase digestion in the DARTS assay in 293T cells. (G) CHX chase analysis of ATF3 protein expression after treatment with PAF in AN3CA and HEC1B cells. (H) WB analysis of ATF3 in ATF3 -overexpressing AN3CA and HEC1B cells. The cells were pretreated with PAF (60 µM) for 24 h and then treated with CHX and MG132 for 24 h. (I) Representative WB images demonstrate the ubiquitination of ATF3 in 293T cells co-transfected with ATF3-Flag, HA-Ub, and plasmids for 24 h. Cellular lysates were collected after 3 h of treatment with PAF, purified with a Flag-tag protein purification kit, and then subjected to WB with anti-HA and anti-ATF3

Article Snippet: SPR assays were performed by TopScience Co., Ltd. (Shanghai, China) to detect the binding between the recombinant ATF3 protein (TMPH-01164, TargetMol) and C16-PAF (74389-68-7, TargetMol), with the ATF3 protein and anti-ATF3 antibody combination used as the positive control.

Techniques: Ubiquitin Proteomics, Binding Assay, SPR Assay, Expressing, Transfection, Purification, FLAG-tag, Protein Purification

Journal: Biology Direct

Article Title: Platelet-activating factor induces ferroptosis by binding to ATF3 and inhibiting the SLC7A11/GPX4 axis to suppress the progression of endometrial carcinoma

doi: 10.1186/s13062-025-00713-z

Figure Lengend Snippet: Schematic overview of the present study. Treatment with PAF attenuates EC cell proliferation by inducing ferroptosis. Mechanistically, PAF interacts with ATF3, enhancing its stability through deubiquitination and consequently suppressing the expression of the ferroptosis-related proteins SLC7A11 and GPX4

Article Snippet: SPR assays were performed by TopScience Co., Ltd. (Shanghai, China) to detect the binding between the recombinant ATF3 protein (TMPH-01164, TargetMol) and C16-PAF (74389-68-7, TargetMol), with the ATF3 protein and anti-ATF3 antibody combination used as the positive control.

Techniques: Expressing

Journal: Journal of Inflammation Research

Article Title: Periodic Mechanical Stress Inhibits the Development of Osteoarthritis via Regulating ATF3-Akt Axis

doi: 10.2147/JIR.S419186

Figure Lengend Snippet: Knockdown of ATF3 alleviates symptoms and pathological knee joint damage in rats with OA. ( A ) Width of the knee joints of rats. ( B ) Knee pain was assessed by weight-bearing test; ( C – E ) Serum levels of TNF-α ( C ), IL-8 ( D ) and IL-6 ( E ) were measured by ELISA (n = 4); ( F ) Histopathological changes of the knee joint from rats (n = 2) were observed by safranin O staining. Scale bar = 100 μm. Error bars are mean ± s.d. * P < 0.05 and ** P < 0.01 vs OA+si-NC group.

Article Snippet: ATF3 antibody, NBP1-85816PEP, 1:1000, Novus Biologicals, Littleton, CO, USA.

Techniques: Knockdown, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Inflammation Research

Article Title: Periodic Mechanical Stress Inhibits the Development of Osteoarthritis via Regulating ATF3-Akt Axis

doi: 10.2147/JIR.S419186

Figure Lengend Snippet: Effects of ATF3 knockdown on extracellular matrix proteins in the articular cartilage of rats with OA. ( A – D ), Expression of collagen II ( A ), aggrecan ( B ) and MMP-13 ( C ) in rat knee joints from rats (n=4) detected by immunohistochemistry and quantified by ImageJ software ( D ). Scale bar = 400 μm. Error bars are mean ± s.d. ** P < 0.01 vs OA+si-NC group.

Article Snippet: ATF3 antibody, NBP1-85816PEP, 1:1000, Novus Biologicals, Littleton, CO, USA.

Techniques: Knockdown, Expressing, Immunohistochemistry, Software

Journal: Journal of Inflammation Research

Article Title: Periodic Mechanical Stress Inhibits the Development of Osteoarthritis via Regulating ATF3-Akt Axis

doi: 10.2147/JIR.S419186

Figure Lengend Snippet: ATF3 regulates the Akt signaling pathway in OA rats. ( A and B ) Western blot was used to detect the protein levels of p-Akt, Akt and ATF3 in the articular cartilage in the Sham and OA groups, Error bars are mean ± s.d. * P < 0.05; ** P < 0.01 vs Sham group; ( C and D ) Western blot was utilized to measure the protein levels of p-Akt, Akt and ATF3 in the articular cartilage in the OA + si-NC and OA + si-ATF3 groups, Error bars are mean ± s.d. ** P < 0.01 vs OA + si-NC group, n = 4.

Article Snippet: ATF3 antibody, NBP1-85816PEP, 1:1000, Novus Biologicals, Littleton, CO, USA.

Techniques: Western Blot

Journal: Journal of Inflammation Research

Article Title: Periodic Mechanical Stress Inhibits the Development of Osteoarthritis via Regulating ATF3-Akt Axis

doi: 10.2147/JIR.S419186

Figure Lengend Snippet: Effects of periodic mechanical stress on inflammation, extracellular matrix proteins and ATF3-Akt axis in osteoarthritic chondrocytes. IL-1β-induced chondrocytes were treated without (group 0) or with PMS under a device height of 1 cm (group 1), 2 cm (group 2), 4 cm (group 4), and 8 cm (group 8). ( A – C ) Levels of TNF-α ( A ), IL-6 ( B ) and IL-8 ( C ) in the supernatant of OA cells measured by ELISA; ( D and E ) Protein expression levels of extracellular matrix proteins (collagen II, aggrecan, and MMP-13) in OA cells detected by Western blot; ( F and G ) Protein expression levels of p-Akt, Akt, and ATF3 in OA cells detected by Western blot. Error bars are mean ± s.d.* P < 0.05; ** P < 0.01 vs Group 0.

Article Snippet: ATF3 antibody, NBP1-85816PEP, 1:1000, Novus Biologicals, Littleton, CO, USA.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Western Blot

Journal: Journal of Inflammation Research

Article Title: Periodic Mechanical Stress Inhibits the Development of Osteoarthritis via Regulating ATF3-Akt Axis

doi: 10.2147/JIR.S419186

Figure Lengend Snippet: Up-regulation of ATF3 reverses the effects of periodic mechanical stress on apoptosis, inflammation and extracellular matrix proteins expression in osteoarthritic chondrocytes. ( A and B ) Apoptotic rate of OA cells was detected by flow cytometry; ( B – D ) Levels of TNF-α ( B ), IL-6 ( C ) and IL-8 ( D ) in OA cells were detected by ELISA; ( E and F ) Protein expression levels of collagen II, aggrecan and MMP-13 in OA cells were detected by Western blot. Error bars are mean ± s.d. * P < 0.05 and ** P < 0.01 vs IL-1β + NC group.

Article Snippet: ATF3 antibody, NBP1-85816PEP, 1:1000, Novus Biologicals, Littleton, CO, USA.

Techniques: Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Journal of Inflammation Research

Article Title: Periodic Mechanical Stress Inhibits the Development of Osteoarthritis via Regulating ATF3-Akt Axis

doi: 10.2147/JIR.S419186

Figure Lengend Snippet: Up-regulation of ATF3 reverses the regulation of the ATF3-Akt axis by periodic mechanical stress in osteoarthritic chondrocytes. ( A and B ) Protein expression levels of p-Akt, ATF3 and Akt in osteoarthritic chondrocytes were detected by Western blot. Error bars are mean ± s.d. ** P < 0.01 vs IL-1β + NC group.

Article Snippet: ATF3 antibody, NBP1-85816PEP, 1:1000, Novus Biologicals, Littleton, CO, USA.

Techniques: Expressing, Western Blot

Journal: Brain structure & function

Article Title: Preservation of KCC2 expression in axotomized abducens motoneurons and its enhancement by VEGF

doi: 10.1007/s00429-023-02635-w

Figure Lengend Snippet: Antibodies used in this study

Article Snippet: ATF3 Used for injured motoneuron identification , Recombinant protein corresponding to aa 1-103 in human ATF3 , Mouse/monoclonal , Novus Clone 1685 NBP2-34489 , AB_2786997 Recognizes the epitope: ASAIVPCLSPPGSL (Manufacturer’s information) Not present in uninjured motoneurons , 1:200.

Techniques: Comparison, Expressing, Recombinant

Journal: Brain structure & function

Article Title: Preservation of KCC2 expression in axotomized abducens motoneurons and its enhancement by VEGF

doi: 10.1007/s00429-023-02635-w

Figure Lengend Snippet: KCC2 immunoreactivity in brainstem oculomotor, trochlear, abducens and facial motoneurons, in control and after axotomy, in the rat. A, C, E, G Confocal images of double immunofluorescence against ChAT (green) and pKCC2 (red) in control oculomotor (A), trochlear (C), abducens (E) and facial (G) motoneurons. B, D, F, H Confocal images of triple immunofluorescence against ChAT (green), KCC2 (red) and ATF3 (white) in the same brainstem nuclei, but after axotomy. Axotomized oculomotor, trochlear and facial motoneurons showed a marked reduction in pKCC2 and ChAT. ATF3 is a general marker of axotomized motoneurons and labels the cell nucleus, as can be observed in axotomized oculomotor (B), trochlear (D) and facial (H) motoneurons (some examples are indicated by white arrows). However, in axotomized abducens motoneurons (F), immunostaining for pKCC2 and ChAT showed a similar appearance to control (E), and ATF3 labeling was absent. I Quantification of KCC2 immunofluorescence in the four nuclei (oculomotor, trochlear, abducens and facial) and in the control and axotomy situation. Asterisks indicate significant (p < 0.001) difference between control and axotomized motoneurons within the same nucleus. Hashtag indicates significant difference (p < 0.001) between axotomized abducens motoneurons and the axotomized motoneurons of the other three nuclei (Two-way ANOVA followed by Holm-Sidak method; n = 35, 36, 33 and 39 for control and n = 33, 35, 44 and 42 for axotomized motoneurons of the oculomotor, trochlear, abducens and facial nuclei, respectively). Data in histograms represent mean ± SEM. Depicted to the right are the Cumming plots of estimated differences after bootstrap resampling with average difference indicated by a dot and 95% CI limit of the distribution by the vertical bars. Oculomotor, trochlear and facial motoneurons all showed significant reduction of 72%, 89% and 87% of the control value, respectively (two-sided permutation t-test p < 0.001 for all comparisons). There were no significant differences when comparing axotomized and control motoneurons in the abducens nucleus. Scale bar = 30 μm in H for A–H

Article Snippet: ATF3 Used for injured motoneuron identification , Recombinant protein corresponding to aa 1-103 in human ATF3 , Mouse/monoclonal , Novus Clone 1685 NBP2-34489 , AB_2786997 Recognizes the epitope: ASAIVPCLSPPGSL (Manufacturer’s information) Not present in uninjured motoneurons , 1:200.

Techniques: Control, Immunofluorescence, Marker, Immunostaining, Labeling

Journal: Brain structure & function

Article Title: Preservation of KCC2 expression in axotomized abducens motoneurons and its enhancement by VEGF

doi: 10.1007/s00429-023-02635-w

Figure Lengend Snippet: KCC2 changes induced by axotomy in cat spinal motoneurons. A–H High magnification single plane confocal images of spinal motoneurons immunostained against ChAT, pKCC2 and ATF3. A–D corresponds to a control motoneuron and E–H to a motoneuron axotomized 21 days previously. Individual immunoreactivities are presented in isolation and then merged (D, H), as indicated in the figure. Axotomized motoneurons expressed ATF3 in the nucleus (arrow in G) and decreased ChAT immunoreactivity (E). They also lacked surface pKCC2 immunoreactivity in the cell body and proximal dendrite surfaces (F). I Axotomized spinal motoneuron immunolabeled for ChAT and ATF3. J KCC2b immunofluorescence of the same axotomized motoneuron as in I illustrating lack of KCC2b labeling in its somatic membrane. K Merge image of I and J. L Comparison of pKCC2 immunofluorescence between control and axotomized spinal motoneurons. Axotomized spinal motoneurons showed a significantly (asterisk) lower level of pKCC2 than control spinal motoneurons (t-test, p ≤ 0.001; n = 96 and 101 control and axotomized motoneurons, respectively). M Swarm dot plots of raw data (individual motoneurons) and differences between control (blue) and axotomy (yellow) shown in Cumming estimation plots. The distribution of differences obtained from bootstrap resampling are shown with the mean difference depicted as a dot and the 95% confidence interval indicated by the ends of the vertical error bars. A 70% significant decrease was detected in axotomized spinal motoneurons (two-sided permutation t-test p < 0.001). N Bar chat illustrating the results of a two-way ANOVA test and Holm-Sidak method comparing the following two factors: experimental condition (control versus axotomy) and type of KCC2 immunoreactivity (pKCC versus KCC2b). Data were gathered from one cat stained in serial sections with pKCC2 and KCC2b (control, n = 29 and 26 motoneurons; axotomy, n = 33 and 27 motoneurons for pKCC2 and KCC2b, respectively). Two-way ANOVA detected significant differences in control vs. axotomized motoneurons (asterisk, p < 0.001), but no difference between pKCC2 and KCC2b (p = 0.099, n.s.), or the interaction between axotomy and type of immunoreactivity (p = 0.334). Post hoc Holm-Sidak pairwise comparisons revealed significant difference between control and injured motoneuron for pKCC2 and KCC2b (#, p < 0.001 for both). O Swarm dot plots of raw data (individual motoneurons) and differences between pKCC2 in control (blue), KCC2b in control (yellow) and pKCC2 after axotomy (green) and KCC2b after axotomy (red) all shown in Cumming estimation plots. The distribution of differences obtained from bootstrap resampling are shown with the mean difference depicted as a dot and the 95% confidence interval indicated by the ends of the vertical error bars. No significant differences were found between KCC2b and pKCC2 in control motoneurons. Axotomized motoneurons showed a 70% significant decrease compared to their respective antibody matched controls (two-sided permutation t-test p < 0.001 in both comparisons). Scale bars = 30 μm in H for A–H; 20 μm in K for I–K

Article Snippet: ATF3 Used for injured motoneuron identification , Recombinant protein corresponding to aa 1-103 in human ATF3 , Mouse/monoclonal , Novus Clone 1685 NBP2-34489 , AB_2786997 Recognizes the epitope: ASAIVPCLSPPGSL (Manufacturer’s information) Not present in uninjured motoneurons , 1:200.

Techniques: Control, Isolation, Immunolabeling, Immunofluorescence, Labeling, Membrane, Comparison, Staining

Journal: eNeuro

Article Title: Removal of the Potassium Chloride Co-Transporter from the Somatodendritic Membrane of Axotomized Motoneurons Is Independent of BDNF/TrkB Signaling But Is Controlled by Neuromuscular Innervation

doi: 10.1523/ENEURO.0172-19.2019

Figure Lengend Snippet: Primary antibodies used for immunohistochemistry (IHC) and Western blottings (WB)

Article Snippet: ATF3 , Recombinant protein corresponding to aa MMLQHPGQVSASEVSASAIVPCLSPPGSLVFEDFANLTPFVKEELRFAINQNKHLCHRMSSALESVTVSDRPLGVSITKAIVAPEEDERKKRRRERNKIAAAKCRNKKKEKTEC , Mouse/ monoclonal , Novus NBP2-34489 , NA , 1:200 , IHC.

Techniques: Immunohistochemistry, Western Blot, Purification, Recombinant, Produced, Phospho-proteomics