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atf3-specific antibodies  (Beyotime)


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    Beyotime atf3-specific antibodies
    Inhibition of the Piezo1 channel alleviates ferroptosis in HUVECs via a Ca 2+ -dependent mechanism. (A) Schematic of in vivo experiment. (B) Wound images during healing and schematic diagram of wound-healing process. (C) Quantitative data of relative wound area to that of day 0 of the 4 groups. (D and E) H&E and Masson staining images on days 3 and 7. (F to H) Representative IHC images of VEGF-A, <t>ATF3,</t> and CaMKII. (I to K) Representative IF images of GPX4, SLC7A11, and α-SMA. (L) Laser Doppler scanned images of subcutaneous vascular flow and blood supply on wounds at days 3 and 7. *** P < 0.001, ns = no significant. All data are performed in triplicate and at least 3 times.
    Atf3 Specific Antibodies, supplied by Beyotime, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/atf3-specific+antibodies/atf3+3+utr++wt+mut/pmc12133029-277-18-21
    Average 90 stars, based on 1 article reviews
    atf3-specific antibodies - by Bioz Stars, 2026-09
    90/100 stars

    Images

    1) Product Images from "Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3"

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3

    Journal: Research

    doi: 10.34133/research.0718

    Inhibition of the Piezo1 channel alleviates ferroptosis in HUVECs via a Ca 2+ -dependent mechanism. (A) Schematic of in vivo experiment. (B) Wound images during healing and schematic diagram of wound-healing process. (C) Quantitative data of relative wound area to that of day 0 of the 4 groups. (D and E) H&E and Masson staining images on days 3 and 7. (F to H) Representative IHC images of VEGF-A, ATF3, and CaMKII. (I to K) Representative IF images of GPX4, SLC7A11, and α-SMA. (L) Laser Doppler scanned images of subcutaneous vascular flow and blood supply on wounds at days 3 and 7. *** P < 0.001, ns = no significant. All data are performed in triplicate and at least 3 times.
    Figure Legend Snippet: Inhibition of the Piezo1 channel alleviates ferroptosis in HUVECs via a Ca 2+ -dependent mechanism. (A) Schematic of in vivo experiment. (B) Wound images during healing and schematic diagram of wound-healing process. (C) Quantitative data of relative wound area to that of day 0 of the 4 groups. (D and E) H&E and Masson staining images on days 3 and 7. (F to H) Representative IHC images of VEGF-A, ATF3, and CaMKII. (I to K) Representative IF images of GPX4, SLC7A11, and α-SMA. (L) Laser Doppler scanned images of subcutaneous vascular flow and blood supply on wounds at days 3 and 7. *** P < 0.001, ns = no significant. All data are performed in triplicate and at least 3 times.

    Techniques Used: Inhibition, In Vivo, Staining

    Blockade of ATF3 protects against ferroptosis in erastin- and Doxo-treated HUVECs. (A) Schematic diagram of RNA sequencing design and sample preparation procedures. (B and C) GSEA shows ferroptosis and lipid metabolism in Doxo group and Doxo + si-Piezo1 group. (D) Venn diagram showing these overlapping genes between 2 clusters (Doxo versus Doxo + si-Piezo1 and Ferroptosis) to obtain the intersection of the 2 DEGs. (E and F) GO and KEGG enrichment analyses showing calcium-mediated signaling, lipid metabolic process, and ferroptosis associated with these DEGs. (G and H) Western blot analysis for n-ATF3 in HUVECs with different treatments. (I and O) Representative IF images and quantification analysis of ATF3. (J to M) Representative images of JC-1 staining, Mito Green, C11-BODIPY staining, and FerroOrange staining in HUVECs with different treatments. (N and P) Western blot analysis for ferroptosis biomarkers ACSL4 and GPX4 in HUVECs with different treatments. (Q) LDH release of HUVECs with different treatments for 24 h. (R) GSH/GSSG ratio was measured in HUVECs with different treatments. *** P < 0.001. All data are performed in triplicate and at least 3 times.
    Figure Legend Snippet: Blockade of ATF3 protects against ferroptosis in erastin- and Doxo-treated HUVECs. (A) Schematic diagram of RNA sequencing design and sample preparation procedures. (B and C) GSEA shows ferroptosis and lipid metabolism in Doxo group and Doxo + si-Piezo1 group. (D) Venn diagram showing these overlapping genes between 2 clusters (Doxo versus Doxo + si-Piezo1 and Ferroptosis) to obtain the intersection of the 2 DEGs. (E and F) GO and KEGG enrichment analyses showing calcium-mediated signaling, lipid metabolic process, and ferroptosis associated with these DEGs. (G and H) Western blot analysis for n-ATF3 in HUVECs with different treatments. (I and O) Representative IF images and quantification analysis of ATF3. (J to M) Representative images of JC-1 staining, Mito Green, C11-BODIPY staining, and FerroOrange staining in HUVECs with different treatments. (N and P) Western blot analysis for ferroptosis biomarkers ACSL4 and GPX4 in HUVECs with different treatments. (Q) LDH release of HUVECs with different treatments for 24 h. (R) GSH/GSSG ratio was measured in HUVECs with different treatments. *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Techniques Used: RNA Sequencing, Sample Prep, Western Blot, Staining

    Activation of Piezo1 by Yoda1 facilitates CaMKII-ATF3 interaction in HUVECs. (A to C) Western blot analysis for n-ATF3, CaMKII, SLC7A11, and GPX4 in HUVECs. (D and E) Representative IF images and quantification analysis of CaMKII and ATF3. (F) Three-dimensional binding structure of CaMKII and ATF3 determined via molecular modeling and docking studies. (G) Close-up view of hydrogen bonding between CaMKII and ATF3. (H) The interaction between CaMKII and ATF3 in HUVECs was analyzed by Co-IP. (I) Schematic diagram of the effects of Doxo and EGTA on the binding between CaMKII and ATF3. *** P < 0.001. All data are performed in triplicate and at least 3 times.
    Figure Legend Snippet: Activation of Piezo1 by Yoda1 facilitates CaMKII-ATF3 interaction in HUVECs. (A to C) Western blot analysis for n-ATF3, CaMKII, SLC7A11, and GPX4 in HUVECs. (D and E) Representative IF images and quantification analysis of CaMKII and ATF3. (F) Three-dimensional binding structure of CaMKII and ATF3 determined via molecular modeling and docking studies. (G) Close-up view of hydrogen bonding between CaMKII and ATF3. (H) The interaction between CaMKII and ATF3 in HUVECs was analyzed by Co-IP. (I) Schematic diagram of the effects of Doxo and EGTA on the binding between CaMKII and ATF3. *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Techniques Used: Activation Assay, Western Blot, Binding Assay, Co-Immunoprecipitation Assay

    ATF3 represses SLC7A11 expression by regulating system Xc − . (A and B) Western blot analysis for ATF3-OE in HUVECs. (C and D) Representative IF images and quantification analysis of ATF3. (E and F) Western blot analysis for n-ATF3 and ferroptosis biomarkers SLC7A11 and GPX4 in HUVECs with different treatments. (G and H) Western blot analysis for ferroptosis biomarkers SLC7A11 and GPX4 in HUVECs with different treatments. (I) GSH/GSSG ratio was measured in HUVECs with different treatments. (J) Predicted ATF3-binding sites in the promoter region of SLC7A11. (K) A schematic illustration depicts the ATF3 motif within the promoter region of the SLC7A11 locus. (L) Luciferase assays were performed in 293T cell. (M) Schematic presentation of a model whereby ATF3 represses SLC7A11 expression to suppress system Xc − , and thereby predisposes cells to a state prone to ferroptosis. (N) The chromatin immunoprecipitation sequencing data previously reported were reanalyzed ( GSM1917770 , ENCSR632DCH_2, GSM803508 , GSM803503 ). *** P < 0.001. All data are performed in triplicate and at least 3 times.
    Figure Legend Snippet: ATF3 represses SLC7A11 expression by regulating system Xc − . (A and B) Western blot analysis for ATF3-OE in HUVECs. (C and D) Representative IF images and quantification analysis of ATF3. (E and F) Western blot analysis for n-ATF3 and ferroptosis biomarkers SLC7A11 and GPX4 in HUVECs with different treatments. (G and H) Western blot analysis for ferroptosis biomarkers SLC7A11 and GPX4 in HUVECs with different treatments. (I) GSH/GSSG ratio was measured in HUVECs with different treatments. (J) Predicted ATF3-binding sites in the promoter region of SLC7A11. (K) A schematic illustration depicts the ATF3 motif within the promoter region of the SLC7A11 locus. (L) Luciferase assays were performed in 293T cell. (M) Schematic presentation of a model whereby ATF3 represses SLC7A11 expression to suppress system Xc − , and thereby predisposes cells to a state prone to ferroptosis. (N) The chromatin immunoprecipitation sequencing data previously reported were reanalyzed ( GSM1917770 , ENCSR632DCH_2, GSM803508 , GSM803503 ). *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Techniques Used: Expressing, Western Blot, Binding Assay, Luciferase, ChIP-sequencing

    The down-expression of ATF3 inhibits ferroptosis through the regulation of the Xc − system. (A) Schematic of in vivo experiment. (B and C) Wound images during healing and schematic diagram of wound-healing process. (D) Quantitative data of relative wound area to that of day 0 of the 4 groups. (E and F) H&E and Masson staining images on days 3 and 7. (G) Representative IHC images of VEGF-A. (H to J) Representative IF images of SLC7A11, α-SMA, and 4-HNE. (K) Laser Doppler scanned images of subcutaneous vascular flow and blood supply on wounds at day 3 and 7. *** P < 0.001. All data are performed in triplicate and at least 3 times.
    Figure Legend Snippet: The down-expression of ATF3 inhibits ferroptosis through the regulation of the Xc − system. (A) Schematic of in vivo experiment. (B and C) Wound images during healing and schematic diagram of wound-healing process. (D) Quantitative data of relative wound area to that of day 0 of the 4 groups. (E and F) H&E and Masson staining images on days 3 and 7. (G) Representative IHC images of VEGF-A. (H to J) Representative IF images of SLC7A11, α-SMA, and 4-HNE. (K) Laser Doppler scanned images of subcutaneous vascular flow and blood supply on wounds at day 3 and 7. *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Techniques Used: Expressing, In Vivo, Staining

    The deficiency of ATF3 promotes wound healing and angiogenesis in aging mice. Piezo1 up-regulated by calcium ion influx triggers ATF3 nuclear translocation via activation of CaMKII, thus aggravating the accumulation of ROS and lipid peroxidation, and eventually leading to ferroptosis in senescent HUVECs.
    Figure Legend Snippet: The deficiency of ATF3 promotes wound healing and angiogenesis in aging mice. Piezo1 up-regulated by calcium ion influx triggers ATF3 nuclear translocation via activation of CaMKII, thus aggravating the accumulation of ROS and lipid peroxidation, and eventually leading to ferroptosis in senescent HUVECs.

    Techniques Used: Translocation Assay, Activation Assay

    Related Articles

    Gentle:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Lysis:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Incubation:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Control:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    SDS Page:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Inhibition:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    In Vivo:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Staining:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    RNA Sequencing:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Sample Prep:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Western Blot:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Activation Assay:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Binding Assay:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Co-Immunoprecipitation Assay:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Expressing:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Luciferase:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    ChIP-sequencing:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Translocation Assay:

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3
    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.



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    reagents antibodies specific for actb q, atf3, fos, and jun  (Santa Cruz Biotechnology)
    90
    Santa Cruz Biotechnology reagents antibodies specific for actb q, atf3, fos, and jun
    Figure 1. Axotomy-induced recombination in peripherally-projecting neurons. A, Validation of an <t>ATF3-specific</t> antibody. The Novus antibody (NBP1-85816) produces a positive signal in nuclei of axotomized sensory neurons in ATF3/ mice, but not ATF3cre/cre mice. Note that the Santa Cruz antibody (C-19) labels neuronal nuclei in in the latter (inset), indicating non-specific staining. B, C, Axotomy induced reporter expression in sensory (DRG), sympathetic (stellate ganglion, SG), and motoneurons 4 d after injury. D, Reporter expression in sensory axons and motoneurons one week after injury. E, Preventing CreERT2 translocation from cytoplasm to nucleus with ICI 182780 reduces recombination in ATF3 cells (by 50%). F, Recombination efficiency 16 d after injury was calculated by expressing the proportion of tracer-filled somata (labeled at the time of injury) that were also reporter (tdtomato)-positive. G, Recombination efficiencies at 4 and 16 d after injury (n 3 for each time point) for DRG and motoneurons. Images in panels A, B, E were taken from whole mounts, those in C, D, F from cryosections.
    Reagents Antibodies Specific For Actb Q, Atf3, Fos, And Jun, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/atf3-specific+antibodies/rabbit+anti+atf3/pm27912076-58-9-13
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    reagents antibodies specific for actb q, atf3, fos, and jun - by Bioz Stars, 2026-09
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    antibodies specific to actin beta (actb), atf3  (Santa Cruz Biotechnology)
    90
    Santa Cruz Biotechnology antibodies specific to actin beta (actb), atf3
    Figure 1. Axotomy-induced recombination in peripherally-projecting neurons. A, Validation of an <t>ATF3-specific</t> antibody. The Novus antibody (NBP1-85816) produces a positive signal in nuclei of axotomized sensory neurons in ATF3/ mice, but not ATF3cre/cre mice. Note that the Santa Cruz antibody (C-19) labels neuronal nuclei in in the latter (inset), indicating non-specific staining. B, C, Axotomy induced reporter expression in sensory (DRG), sympathetic (stellate ganglion, SG), and motoneurons 4 d after injury. D, Reporter expression in sensory axons and motoneurons one week after injury. E, Preventing CreERT2 translocation from cytoplasm to nucleus with ICI 182780 reduces recombination in ATF3 cells (by 50%). F, Recombination efficiency 16 d after injury was calculated by expressing the proportion of tracer-filled somata (labeled at the time of injury) that were also reporter (tdtomato)-positive. G, Recombination efficiencies at 4 and 16 d after injury (n 3 for each time point) for DRG and motoneurons. Images in panels A, B, E were taken from whole mounts, those in C, D, F from cryosections.
    Antibodies Specific To Actin Beta (Actb), Atf3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/atf3-specific+antibodies/rabbit+anti+atf3/pm28185985-65-6-9
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    Image Search Results


    Inhibition of the Piezo1 channel alleviates ferroptosis in HUVECs via a Ca 2+ -dependent mechanism. (A) Schematic of in vivo experiment. (B) Wound images during healing and schematic diagram of wound-healing process. (C) Quantitative data of relative wound area to that of day 0 of the 4 groups. (D and E) H&E and Masson staining images on days 3 and 7. (F to H) Representative IHC images of VEGF-A, ATF3, and CaMKII. (I to K) Representative IF images of GPX4, SLC7A11, and α-SMA. (L) Laser Doppler scanned images of subcutaneous vascular flow and blood supply on wounds at days 3 and 7. *** P < 0.001, ns = no significant. All data are performed in triplicate and at least 3 times.

    Journal: Research

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3

    doi: 10.34133/research.0718

    Figure Lengend Snippet: Inhibition of the Piezo1 channel alleviates ferroptosis in HUVECs via a Ca 2+ -dependent mechanism. (A) Schematic of in vivo experiment. (B) Wound images during healing and schematic diagram of wound-healing process. (C) Quantitative data of relative wound area to that of day 0 of the 4 groups. (D and E) H&E and Masson staining images on days 3 and 7. (F to H) Representative IHC images of VEGF-A, ATF3, and CaMKII. (I to K) Representative IF images of GPX4, SLC7A11, and α-SMA. (L) Laser Doppler scanned images of subcutaneous vascular flow and blood supply on wounds at days 3 and 7. *** P < 0.001, ns = no significant. All data are performed in triplicate and at least 3 times.

    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Techniques: Inhibition, In Vivo, Staining

    Blockade of ATF3 protects against ferroptosis in erastin- and Doxo-treated HUVECs. (A) Schematic diagram of RNA sequencing design and sample preparation procedures. (B and C) GSEA shows ferroptosis and lipid metabolism in Doxo group and Doxo + si-Piezo1 group. (D) Venn diagram showing these overlapping genes between 2 clusters (Doxo versus Doxo + si-Piezo1 and Ferroptosis) to obtain the intersection of the 2 DEGs. (E and F) GO and KEGG enrichment analyses showing calcium-mediated signaling, lipid metabolic process, and ferroptosis associated with these DEGs. (G and H) Western blot analysis for n-ATF3 in HUVECs with different treatments. (I and O) Representative IF images and quantification analysis of ATF3. (J to M) Representative images of JC-1 staining, Mito Green, C11-BODIPY staining, and FerroOrange staining in HUVECs with different treatments. (N and P) Western blot analysis for ferroptosis biomarkers ACSL4 and GPX4 in HUVECs with different treatments. (Q) LDH release of HUVECs with different treatments for 24 h. (R) GSH/GSSG ratio was measured in HUVECs with different treatments. *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Journal: Research

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3

    doi: 10.34133/research.0718

    Figure Lengend Snippet: Blockade of ATF3 protects against ferroptosis in erastin- and Doxo-treated HUVECs. (A) Schematic diagram of RNA sequencing design and sample preparation procedures. (B and C) GSEA shows ferroptosis and lipid metabolism in Doxo group and Doxo + si-Piezo1 group. (D) Venn diagram showing these overlapping genes between 2 clusters (Doxo versus Doxo + si-Piezo1 and Ferroptosis) to obtain the intersection of the 2 DEGs. (E and F) GO and KEGG enrichment analyses showing calcium-mediated signaling, lipid metabolic process, and ferroptosis associated with these DEGs. (G and H) Western blot analysis for n-ATF3 in HUVECs with different treatments. (I and O) Representative IF images and quantification analysis of ATF3. (J to M) Representative images of JC-1 staining, Mito Green, C11-BODIPY staining, and FerroOrange staining in HUVECs with different treatments. (N and P) Western blot analysis for ferroptosis biomarkers ACSL4 and GPX4 in HUVECs with different treatments. (Q) LDH release of HUVECs with different treatments for 24 h. (R) GSH/GSSG ratio was measured in HUVECs with different treatments. *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Techniques: RNA Sequencing, Sample Prep, Western Blot, Staining

    Activation of Piezo1 by Yoda1 facilitates CaMKII-ATF3 interaction in HUVECs. (A to C) Western blot analysis for n-ATF3, CaMKII, SLC7A11, and GPX4 in HUVECs. (D and E) Representative IF images and quantification analysis of CaMKII and ATF3. (F) Three-dimensional binding structure of CaMKII and ATF3 determined via molecular modeling and docking studies. (G) Close-up view of hydrogen bonding between CaMKII and ATF3. (H) The interaction between CaMKII and ATF3 in HUVECs was analyzed by Co-IP. (I) Schematic diagram of the effects of Doxo and EGTA on the binding between CaMKII and ATF3. *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Journal: Research

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3

    doi: 10.34133/research.0718

    Figure Lengend Snippet: Activation of Piezo1 by Yoda1 facilitates CaMKII-ATF3 interaction in HUVECs. (A to C) Western blot analysis for n-ATF3, CaMKII, SLC7A11, and GPX4 in HUVECs. (D and E) Representative IF images and quantification analysis of CaMKII and ATF3. (F) Three-dimensional binding structure of CaMKII and ATF3 determined via molecular modeling and docking studies. (G) Close-up view of hydrogen bonding between CaMKII and ATF3. (H) The interaction between CaMKII and ATF3 in HUVECs was analyzed by Co-IP. (I) Schematic diagram of the effects of Doxo and EGTA on the binding between CaMKII and ATF3. *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Techniques: Activation Assay, Western Blot, Binding Assay, Co-Immunoprecipitation Assay

    ATF3 represses SLC7A11 expression by regulating system Xc − . (A and B) Western blot analysis for ATF3-OE in HUVECs. (C and D) Representative IF images and quantification analysis of ATF3. (E and F) Western blot analysis for n-ATF3 and ferroptosis biomarkers SLC7A11 and GPX4 in HUVECs with different treatments. (G and H) Western blot analysis for ferroptosis biomarkers SLC7A11 and GPX4 in HUVECs with different treatments. (I) GSH/GSSG ratio was measured in HUVECs with different treatments. (J) Predicted ATF3-binding sites in the promoter region of SLC7A11. (K) A schematic illustration depicts the ATF3 motif within the promoter region of the SLC7A11 locus. (L) Luciferase assays were performed in 293T cell. (M) Schematic presentation of a model whereby ATF3 represses SLC7A11 expression to suppress system Xc − , and thereby predisposes cells to a state prone to ferroptosis. (N) The chromatin immunoprecipitation sequencing data previously reported were reanalyzed ( GSM1917770 , ENCSR632DCH_2, GSM803508 , GSM803503 ). *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Journal: Research

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3

    doi: 10.34133/research.0718

    Figure Lengend Snippet: ATF3 represses SLC7A11 expression by regulating system Xc − . (A and B) Western blot analysis for ATF3-OE in HUVECs. (C and D) Representative IF images and quantification analysis of ATF3. (E and F) Western blot analysis for n-ATF3 and ferroptosis biomarkers SLC7A11 and GPX4 in HUVECs with different treatments. (G and H) Western blot analysis for ferroptosis biomarkers SLC7A11 and GPX4 in HUVECs with different treatments. (I) GSH/GSSG ratio was measured in HUVECs with different treatments. (J) Predicted ATF3-binding sites in the promoter region of SLC7A11. (K) A schematic illustration depicts the ATF3 motif within the promoter region of the SLC7A11 locus. (L) Luciferase assays were performed in 293T cell. (M) Schematic presentation of a model whereby ATF3 represses SLC7A11 expression to suppress system Xc − , and thereby predisposes cells to a state prone to ferroptosis. (N) The chromatin immunoprecipitation sequencing data previously reported were reanalyzed ( GSM1917770 , ENCSR632DCH_2, GSM803508 , GSM803503 ). *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Techniques: Expressing, Western Blot, Binding Assay, Luciferase, ChIP-sequencing

    The down-expression of ATF3 inhibits ferroptosis through the regulation of the Xc − system. (A) Schematic of in vivo experiment. (B and C) Wound images during healing and schematic diagram of wound-healing process. (D) Quantitative data of relative wound area to that of day 0 of the 4 groups. (E and F) H&E and Masson staining images on days 3 and 7. (G) Representative IHC images of VEGF-A. (H to J) Representative IF images of SLC7A11, α-SMA, and 4-HNE. (K) Laser Doppler scanned images of subcutaneous vascular flow and blood supply on wounds at day 3 and 7. *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Journal: Research

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3

    doi: 10.34133/research.0718

    Figure Lengend Snippet: The down-expression of ATF3 inhibits ferroptosis through the regulation of the Xc − system. (A) Schematic of in vivo experiment. (B and C) Wound images during healing and schematic diagram of wound-healing process. (D) Quantitative data of relative wound area to that of day 0 of the 4 groups. (E and F) H&E and Masson staining images on days 3 and 7. (G) Representative IHC images of VEGF-A. (H to J) Representative IF images of SLC7A11, α-SMA, and 4-HNE. (K) Laser Doppler scanned images of subcutaneous vascular flow and blood supply on wounds at day 3 and 7. *** P < 0.001. All data are performed in triplicate and at least 3 times.

    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Techniques: Expressing, In Vivo, Staining

    The deficiency of ATF3 promotes wound healing and angiogenesis in aging mice. Piezo1 up-regulated by calcium ion influx triggers ATF3 nuclear translocation via activation of CaMKII, thus aggravating the accumulation of ROS and lipid peroxidation, and eventually leading to ferroptosis in senescent HUVECs.

    Journal: Research

    Article Title: Piezo1-Mediated Ferroptosis Delays Wound Healing in Aging Mice by Regulating the Transcriptional Activity of SLC7A11 through Activating Transcription Factor 3

    doi: 10.34133/research.0718

    Figure Lengend Snippet: The deficiency of ATF3 promotes wound healing and angiogenesis in aging mice. Piezo1 up-regulated by calcium ion influx triggers ATF3 nuclear translocation via activation of CaMKII, thus aggravating the accumulation of ROS and lipid peroxidation, and eventually leading to ferroptosis in senescent HUVECs.

    Article Snippet: Cells were fixed and lysed in a gentle lysis buffer, and 90% of the lysate was incubated with ATF3-specific antibodies (from Beyotime, catalog no. P2171, China) and an immunoglobulin G (IgG) control (same supplier) for 24 h. Complexes were then pelleted, washed 3 times with lysis buffer, and denatured before being loaded onto a denaturing SDS-PAGE gel for analysis.

    Techniques: Translocation Assay, Activation Assay

    Axotomy-induced recombination in peripherally-projecting neurons. A , Validation of an ATF3-specific antibody. The Novus antibody (NBP1-85816) produces a positive signal in nuclei of axotomized sensory neurons in ATF3 +/+ mice, but not ATF3 cre/cre mice. Note that the Santa Cruz antibody (C-19) labels neuronal nuclei in in the latter (inset), indicating non-specific staining. B , C , Axotomy induced reporter expression in sensory (DRG), sympathetic (stellate ganglion, SG), and motoneurons 4 d after injury. D , Reporter expression in sensory axons and motoneurons one week after injury. E , Preventing CreERT2 translocation from cytoplasm to nucleus with ICI 182780 reduces recombination in ATF3 + cells (by ∼50%). F , Recombination efficiency 16 d after injury was calculated by expressing the proportion of tracer-filled somata (labeled at the time of injury) that were also reporter (tdtomato)-positive. G , Recombination efficiencies at 4 and 16 d after injury ( n = 3 for each time point) for DRG and motoneurons. Images in panels A , B , E were taken from whole mounts, those in C , D , F from cryosections.

    Journal: eNeuro

    Article Title: An ATF3-CreERT2 Knock-In Mouse for Axotomy-Induced Genetic Editing: Proof of Principle

    doi: 10.1523/ENEURO.0025-19.2019

    Figure Lengend Snippet: Axotomy-induced recombination in peripherally-projecting neurons. A , Validation of an ATF3-specific antibody. The Novus antibody (NBP1-85816) produces a positive signal in nuclei of axotomized sensory neurons in ATF3 +/+ mice, but not ATF3 cre/cre mice. Note that the Santa Cruz antibody (C-19) labels neuronal nuclei in in the latter (inset), indicating non-specific staining. B , C , Axotomy induced reporter expression in sensory (DRG), sympathetic (stellate ganglion, SG), and motoneurons 4 d after injury. D , Reporter expression in sensory axons and motoneurons one week after injury. E , Preventing CreERT2 translocation from cytoplasm to nucleus with ICI 182780 reduces recombination in ATF3 + cells (by ∼50%). F , Recombination efficiency 16 d after injury was calculated by expressing the proportion of tracer-filled somata (labeled at the time of injury) that were also reporter (tdtomato)-positive. G , Recombination efficiencies at 4 and 16 d after injury ( n = 3 for each time point) for DRG and motoneurons. Images in panels A , B , E were taken from whole mounts, those in C , D , F from cryosections.

    Article Snippet: Using a homozygous knock-in (i.e., null mutant), we tested the validity of two reported ATF3-specific polyclonal antibodies: Novus (NBP 1-85816) and Santa Cruz (C-19).

    Techniques: Biomarker Discovery, Staining, Expressing, Translocation Assay, Labeling

    Axotomy-induced PTEN deletion and anatomic regeneration following sciatic nerve crush. A , B , Axonal regeneration 2 d subsequent to sciatic nerve crush (dotted line) was significantly enhanced in ATF3 +/cre :PTEN fl/fl mice. In C , bar graphs represent the cumulative density of SCG10 immunoreactivity from 0 μm (the distal extent of the crush site) to 2000 μm; n = 6 for both groups and cumulative densities were compared using an unpaired t test. Scale bars: 500 μm.

    Journal: eNeuro

    Article Title: An ATF3-CreERT2 Knock-In Mouse for Axotomy-Induced Genetic Editing: Proof of Principle

    doi: 10.1523/ENEURO.0025-19.2019

    Figure Lengend Snippet: Axotomy-induced PTEN deletion and anatomic regeneration following sciatic nerve crush. A , B , Axonal regeneration 2 d subsequent to sciatic nerve crush (dotted line) was significantly enhanced in ATF3 +/cre :PTEN fl/fl mice. In C , bar graphs represent the cumulative density of SCG10 immunoreactivity from 0 μm (the distal extent of the crush site) to 2000 μm; n = 6 for both groups and cumulative densities were compared using an unpaired t test. Scale bars: 500 μm.

    Article Snippet: Using a homozygous knock-in (i.e., null mutant), we tested the validity of two reported ATF3-specific polyclonal antibodies: Novus (NBP 1-85816) and Santa Cruz (C-19).

    Techniques:

    Figure 1. Axotomy-induced recombination in peripherally-projecting neurons. A, Validation of an ATF3-specific antibody. The Novus antibody (NBP1-85816) produces a positive signal in nuclei of axotomized sensory neurons in ATF3/ mice, but not ATF3cre/cre mice. Note that the Santa Cruz antibody (C-19) labels neuronal nuclei in in the latter (inset), indicating non-specific staining. B, C, Axotomy induced reporter expression in sensory (DRG), sympathetic (stellate ganglion, SG), and motoneurons 4 d after injury. D, Reporter expression in sensory axons and motoneurons one week after injury. E, Preventing CreERT2 translocation from cytoplasm to nucleus with ICI 182780 reduces recombination in ATF3 cells (by 50%). F, Recombination efficiency 16 d after injury was calculated by expressing the proportion of tracer-filled somata (labeled at the time of injury) that were also reporter (tdtomato)-positive. G, Recombination efficiencies at 4 and 16 d after injury (n 3 for each time point) for DRG and motoneurons. Images in panels A, B, E were taken from whole mounts, those in C, D, F from cryosections.

    Journal: eneuro

    Article Title: An ATF3-CreERT2 Knock-In Mouse for Axotomy-Induced Genetic Editing: Proof of Principle

    doi: 10.1523/eneuro.0025-19.2019

    Figure Lengend Snippet: Figure 1. Axotomy-induced recombination in peripherally-projecting neurons. A, Validation of an ATF3-specific antibody. The Novus antibody (NBP1-85816) produces a positive signal in nuclei of axotomized sensory neurons in ATF3/ mice, but not ATF3cre/cre mice. Note that the Santa Cruz antibody (C-19) labels neuronal nuclei in in the latter (inset), indicating non-specific staining. B, C, Axotomy induced reporter expression in sensory (DRG), sympathetic (stellate ganglion, SG), and motoneurons 4 d after injury. D, Reporter expression in sensory axons and motoneurons one week after injury. E, Preventing CreERT2 translocation from cytoplasm to nucleus with ICI 182780 reduces recombination in ATF3 cells (by 50%). F, Recombination efficiency 16 d after injury was calculated by expressing the proportion of tracer-filled somata (labeled at the time of injury) that were also reporter (tdtomato)-positive. G, Recombination efficiencies at 4 and 16 d after injury (n 3 for each time point) for DRG and motoneurons. Images in panels A, B, E were taken from whole mounts, those in C, D, F from cryosections.

    Article Snippet: Using a homozygous knock-in (i.e., null mutant), we tested the validity of two reported ATF3-specific polyclonal antibodies: Novus (NBP 1-85816) and Santa Cruz (C-19).

    Techniques: Biomarker Discovery, Staining, Expressing, Translocation Assay, Labeling

    Figure 2. Axotomy does not induce ATF3 in Schwann cells. A, Cryosections from injured DRG (inset) and distal sciatic nerve from the same mouse processed for ATF3 immunohistochemistry (Novus NBP1-85816). B, Punctate staining in the nerve proved to be non-specific fluorescence of leukocytes (note non-nuclear signal in the absence of primary antibody). C, In intact sciatic nerves, cells morphologically identical to Remak cells had at some point undergone recombination. D, Following injury, their numbers increased. E, This was attributable to their proliferation in the injured nerve (as opposed to ATF3 induction and subsequent recombination). C=, D=, Magnification of areas outlined in C and D demonstrate the spindle shaped morphology characteristic of Remak cells.

    Journal: eneuro

    Article Title: An ATF3-CreERT2 Knock-In Mouse for Axotomy-Induced Genetic Editing: Proof of Principle

    doi: 10.1523/eneuro.0025-19.2019

    Figure Lengend Snippet: Figure 2. Axotomy does not induce ATF3 in Schwann cells. A, Cryosections from injured DRG (inset) and distal sciatic nerve from the same mouse processed for ATF3 immunohistochemistry (Novus NBP1-85816). B, Punctate staining in the nerve proved to be non-specific fluorescence of leukocytes (note non-nuclear signal in the absence of primary antibody). C, In intact sciatic nerves, cells morphologically identical to Remak cells had at some point undergone recombination. D, Following injury, their numbers increased. E, This was attributable to their proliferation in the injured nerve (as opposed to ATF3 induction and subsequent recombination). C=, D=, Magnification of areas outlined in C and D demonstrate the spindle shaped morphology characteristic of Remak cells.

    Article Snippet: Using a homozygous knock-in (i.e., null mutant), we tested the validity of two reported ATF3-specific polyclonal antibodies: Novus (NBP 1-85816) and Santa Cruz (C-19).

    Techniques: Immunohistochemistry, Staining, Fluorescence

    Figure 3. Loss of ATF3 function delays functional recovery following sciatic nerve crush. A, B, The rate of functional recovery

    Journal: eneuro

    Article Title: An ATF3-CreERT2 Knock-In Mouse for Axotomy-Induced Genetic Editing: Proof of Principle

    doi: 10.1523/eneuro.0025-19.2019

    Figure Lengend Snippet: Figure 3. Loss of ATF3 function delays functional recovery following sciatic nerve crush. A, B, The rate of functional recovery

    Article Snippet: Using a homozygous knock-in (i.e., null mutant), we tested the validity of two reported ATF3-specific polyclonal antibodies: Novus (NBP 1-85816) and Santa Cruz (C-19).

    Techniques: Functional Assay

    Figure 4. Loss of ATF3 function modestly reduces axonal regeneration following sciatic nerve crush. A, B, There was no difference in axonal regeneration 2 d following injury between ATF3/, ATF3/cre, and ATF3cre/cre mice (n 7, n 6, and n 6, respectively, groups were compared with a one-way ANOVA on cumulative densities). C, D, ATF3cre/cre mice exhibited significantly diminished axonal regeneration 3 d following injury 2–4 mm distal to the injury compared to ATF3/ mice (n 5 for both groups, one-way

    Journal: eneuro

    Article Title: An ATF3-CreERT2 Knock-In Mouse for Axotomy-Induced Genetic Editing: Proof of Principle

    doi: 10.1523/eneuro.0025-19.2019

    Figure Lengend Snippet: Figure 4. Loss of ATF3 function modestly reduces axonal regeneration following sciatic nerve crush. A, B, There was no difference in axonal regeneration 2 d following injury between ATF3/, ATF3/cre, and ATF3cre/cre mice (n 7, n 6, and n 6, respectively, groups were compared with a one-way ANOVA on cumulative densities). C, D, ATF3cre/cre mice exhibited significantly diminished axonal regeneration 3 d following injury 2–4 mm distal to the injury compared to ATF3/ mice (n 5 for both groups, one-way

    Article Snippet: Using a homozygous knock-in (i.e., null mutant), we tested the validity of two reported ATF3-specific polyclonal antibodies: Novus (NBP 1-85816) and Santa Cruz (C-19).

    Techniques:

    Figure 5. Axotomy-induced PTEN deletion in sensory and motoneurons. A, B, Axotomy results in significant loss of PTEN expression in the DRG. Arrows in A indicate small PTEN-positive DRG neurons. The PTEN antibody results in high background staining in all animals, to which PTEN immunoreactivity is reduced in axotomized DRGs of ATF3/cre:PTENfl/fl mice (B; one-way ANOVA followed by Dunnett’s multiple comparison test, n 5 for both groups). C, PTEN immunoreactivity is weak in all motoneurons, rendering difficult confirmation of axotomy-induced knock-down. However, ventral root (VR; large arrow) axons close to the ventral root exit zone were intensely immunopositive, single arrows indicate axons that were both tdtomato and PTEN positive, whereas double arrows indicate recombination without PTEN immunoreactivity. In sections of ventral roots, we were able to demonstrate a significant decrease in PTEN immunoreactivity in tdtomato-positive axons (D; Kolmogorov–Smirnov goodness of fit test), n 4 and n 3 for ATF3/cre:PTEN/ and ATF3/cre:PTENfl/fl, respectively. Scale bars: 50 m (A), 100 m (C), 10 m (E).

    Journal: eneuro

    Article Title: An ATF3-CreERT2 Knock-In Mouse for Axotomy-Induced Genetic Editing: Proof of Principle

    doi: 10.1523/eneuro.0025-19.2019

    Figure Lengend Snippet: Figure 5. Axotomy-induced PTEN deletion in sensory and motoneurons. A, B, Axotomy results in significant loss of PTEN expression in the DRG. Arrows in A indicate small PTEN-positive DRG neurons. The PTEN antibody results in high background staining in all animals, to which PTEN immunoreactivity is reduced in axotomized DRGs of ATF3/cre:PTENfl/fl mice (B; one-way ANOVA followed by Dunnett’s multiple comparison test, n 5 for both groups). C, PTEN immunoreactivity is weak in all motoneurons, rendering difficult confirmation of axotomy-induced knock-down. However, ventral root (VR; large arrow) axons close to the ventral root exit zone were intensely immunopositive, single arrows indicate axons that were both tdtomato and PTEN positive, whereas double arrows indicate recombination without PTEN immunoreactivity. In sections of ventral roots, we were able to demonstrate a significant decrease in PTEN immunoreactivity in tdtomato-positive axons (D; Kolmogorov–Smirnov goodness of fit test), n 4 and n 3 for ATF3/cre:PTEN/ and ATF3/cre:PTENfl/fl, respectively. Scale bars: 50 m (A), 100 m (C), 10 m (E).

    Article Snippet: Using a homozygous knock-in (i.e., null mutant), we tested the validity of two reported ATF3-specific polyclonal antibodies: Novus (NBP 1-85816) and Santa Cruz (C-19).

    Techniques: Expressing, Staining, Comparison, Knockdown

    Figure 7. Axotomy-induced PTEN deletion and anatomic regeneration following sciatic nerve crush. A, B, Axonal regeneration 2 d subsequent to sciatic nerve crush (dotted line) was significantly enhanced in ATF3/cre:PTENfl/fl mice. In C, bar graphs represent the cumulative density of SCG10 immunoreactivity from 0 m (the distal extent of the crush site) to 2000 m; n 6 for both groups and cumulative densities were compared using an unpaired t test. Scale bars: 500 m.

    Journal: eneuro

    Article Title: An ATF3-CreERT2 Knock-In Mouse for Axotomy-Induced Genetic Editing: Proof of Principle

    doi: 10.1523/eneuro.0025-19.2019

    Figure Lengend Snippet: Figure 7. Axotomy-induced PTEN deletion and anatomic regeneration following sciatic nerve crush. A, B, Axonal regeneration 2 d subsequent to sciatic nerve crush (dotted line) was significantly enhanced in ATF3/cre:PTENfl/fl mice. In C, bar graphs represent the cumulative density of SCG10 immunoreactivity from 0 m (the distal extent of the crush site) to 2000 m; n 6 for both groups and cumulative densities were compared using an unpaired t test. Scale bars: 500 m.

    Article Snippet: Using a homozygous knock-in (i.e., null mutant), we tested the validity of two reported ATF3-specific polyclonal antibodies: Novus (NBP 1-85816) and Santa Cruz (C-19).

    Techniques:

    Figure 9. Hemisection-induced recombination in supraspinal neurons one week after injury. A, Paraventricular hypothalamic nucleus, descending part. B, Red nucleus. C, Vestibulospinal nucleus (the genu of the facial nerve is indicated by VII). D, Reticulospinal neurons (RtS) and raphespinal neurons (arrow), and the rubrospinal tract (RST). E, Reporter-expressing axons in the medial longitudinal fasciculus (conveying descending projections of reticulospinal and vestibulospinal axons). F, Examples of ATF3-positive, tdtomato- positive and tdtomato-negative neurons in the paraventricular hypothalamic nucleus (PHN), red nucleus (RN), vestibulospinal nucleus (VSN), and reticulospinal neurons (RtS).

    Journal: eneuro

    Article Title: An ATF3-CreERT2 Knock-In Mouse for Axotomy-Induced Genetic Editing: Proof of Principle

    doi: 10.1523/eneuro.0025-19.2019

    Figure Lengend Snippet: Figure 9. Hemisection-induced recombination in supraspinal neurons one week after injury. A, Paraventricular hypothalamic nucleus, descending part. B, Red nucleus. C, Vestibulospinal nucleus (the genu of the facial nerve is indicated by VII). D, Reticulospinal neurons (RtS) and raphespinal neurons (arrow), and the rubrospinal tract (RST). E, Reporter-expressing axons in the medial longitudinal fasciculus (conveying descending projections of reticulospinal and vestibulospinal axons). F, Examples of ATF3-positive, tdtomato- positive and tdtomato-negative neurons in the paraventricular hypothalamic nucleus (PHN), red nucleus (RN), vestibulospinal nucleus (VSN), and reticulospinal neurons (RtS).

    Article Snippet: Using a homozygous knock-in (i.e., null mutant), we tested the validity of two reported ATF3-specific polyclonal antibodies: Novus (NBP 1-85816) and Santa Cruz (C-19).

    Techniques: Expressing