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p atf2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p atf2
    Effect of transient SUMOylation inhibition in pre-adipocytes on cAMP-PKA-p38 signaling and adaptive thermogenesis in mature adipocytes. ( A ) Heatmap showing the stable upregulation of most genes involved in adaptive thermogenesis and cAMP-PKA-p38 signaling 22 days after adipogenic induction. Z -scores were calculated and plotted in GraphPad Prism. Treatments of pre-adipocytes were performed as shown in Fig. . ( B ) Western blot analysis of PKA substrates phosphorylation, assessed using a pan-PKA-target antibody, 22 days after adipogenic induction. TBP was used as a loading control. The asterisk (*) indicates substrates significantly affected by TAK-981 and/or rosiglitazone treatment. ( C ) Quantification of PKA substrate signals from panel (B). The signal for PKA substrates and TBP was quantified using Fiji software, with PKA substrate signals normalized to TBP. Error bars represent the standard deviation of four independent experiments. Student’s t -test was used to calculate P -values. ( D ) Western blot analysis of p-CREB, p-ATF1, p38, p-p38, and <t>p-ATF2,</t> 22 days after adipogenic induction. TBP was used as a loading control. ( E ) Quantification of p-CREB/ATF-1 signals from panel (D). Signals were normalized to TBP. Error bars represent the standard deviation of three independent experiments. Student’s t -test was used to calculate P -values. ( F ) Western blot analysis of p-CREB in the absence or presence of the PKA inhibitor RP-8-CPT-cAMPS. TBP was used as a loading control. Quantification of p-CREB, normalized to TBP, is showed in the lower panel. Error bars represent the standard deviation of two independent experiments. Quantification of p-p38 ( G ) and <t>p-ATF2</t> ( H ) from experiment in panel (D). Signals were normalized to TBP for p-ATF2 and to p38 for p-p38. Error bars represent the standard deviation of three independent experiments. Student’s t -test was used to calculate P -values.
    P Atf2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Transient SUMOylation inhibition in human pre-adipocytes stably imprints a transcriptional beiging fate"

    Article Title: Transient SUMOylation inhibition in human pre-adipocytes stably imprints a transcriptional beiging fate

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkag232

    Effect of transient SUMOylation inhibition in pre-adipocytes on cAMP-PKA-p38 signaling and adaptive thermogenesis in mature adipocytes. ( A ) Heatmap showing the stable upregulation of most genes involved in adaptive thermogenesis and cAMP-PKA-p38 signaling 22 days after adipogenic induction. Z -scores were calculated and plotted in GraphPad Prism. Treatments of pre-adipocytes were performed as shown in Fig. . ( B ) Western blot analysis of PKA substrates phosphorylation, assessed using a pan-PKA-target antibody, 22 days after adipogenic induction. TBP was used as a loading control. The asterisk (*) indicates substrates significantly affected by TAK-981 and/or rosiglitazone treatment. ( C ) Quantification of PKA substrate signals from panel (B). The signal for PKA substrates and TBP was quantified using Fiji software, with PKA substrate signals normalized to TBP. Error bars represent the standard deviation of four independent experiments. Student’s t -test was used to calculate P -values. ( D ) Western blot analysis of p-CREB, p-ATF1, p38, p-p38, and p-ATF2, 22 days after adipogenic induction. TBP was used as a loading control. ( E ) Quantification of p-CREB/ATF-1 signals from panel (D). Signals were normalized to TBP. Error bars represent the standard deviation of three independent experiments. Student’s t -test was used to calculate P -values. ( F ) Western blot analysis of p-CREB in the absence or presence of the PKA inhibitor RP-8-CPT-cAMPS. TBP was used as a loading control. Quantification of p-CREB, normalized to TBP, is showed in the lower panel. Error bars represent the standard deviation of two independent experiments. Quantification of p-p38 ( G ) and p-ATF2 ( H ) from experiment in panel (D). Signals were normalized to TBP for p-ATF2 and to p38 for p-p38. Error bars represent the standard deviation of three independent experiments. Student’s t -test was used to calculate P -values.
    Figure Legend Snippet: Effect of transient SUMOylation inhibition in pre-adipocytes on cAMP-PKA-p38 signaling and adaptive thermogenesis in mature adipocytes. ( A ) Heatmap showing the stable upregulation of most genes involved in adaptive thermogenesis and cAMP-PKA-p38 signaling 22 days after adipogenic induction. Z -scores were calculated and plotted in GraphPad Prism. Treatments of pre-adipocytes were performed as shown in Fig. . ( B ) Western blot analysis of PKA substrates phosphorylation, assessed using a pan-PKA-target antibody, 22 days after adipogenic induction. TBP was used as a loading control. The asterisk (*) indicates substrates significantly affected by TAK-981 and/or rosiglitazone treatment. ( C ) Quantification of PKA substrate signals from panel (B). The signal for PKA substrates and TBP was quantified using Fiji software, with PKA substrate signals normalized to TBP. Error bars represent the standard deviation of four independent experiments. Student’s t -test was used to calculate P -values. ( D ) Western blot analysis of p-CREB, p-ATF1, p38, p-p38, and p-ATF2, 22 days after adipogenic induction. TBP was used as a loading control. ( E ) Quantification of p-CREB/ATF-1 signals from panel (D). Signals were normalized to TBP. Error bars represent the standard deviation of three independent experiments. Student’s t -test was used to calculate P -values. ( F ) Western blot analysis of p-CREB in the absence or presence of the PKA inhibitor RP-8-CPT-cAMPS. TBP was used as a loading control. Quantification of p-CREB, normalized to TBP, is showed in the lower panel. Error bars represent the standard deviation of two independent experiments. Quantification of p-p38 ( G ) and p-ATF2 ( H ) from experiment in panel (D). Signals were normalized to TBP for p-ATF2 and to p38 for p-p38. Error bars represent the standard deviation of three independent experiments. Student’s t -test was used to calculate P -values.

    Techniques Used: Inhibition, Western Blot, Phospho-proteomics, Control, Software, Standard Deviation

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    Article Snippet: .. Western Blot Analysis Monoclonal antibodies specific for phospho-EGFR (Tyr1068; Tyr1148), Akt, phospho-Akt (Thr308), phospho-Akt (Ser473), phospho-PDK1 (Ser241), phospho-GSK-3a/b (Ser21/9), phosphoCyclin D1 (Thr286), phospho-p44/42 MAPK (Thr202/Thr204), p44/42 MAPK, PTEN, phospho-PTEN (Ser380), phosphomTOR (Ser2448), phospho-p70S6K (Thr389), phospho-p38 MAPK (Thr180/Tyr182), phospho-ATF-2 (Thr71), phospho-STAT3 (Ser727), phospho-STAT3 (Tyr705), phospho-STAT5 (Tyr694), phospho-c-Fos (Ser32) and Cox2 were purchased from Cell Signaling Technology. .. Antibodies against surfactant protein C (SPC), Clara cell 10 (CC10) protein and actin were purchased from Santa Cruz Biotechnology.

    Article Title: Sodium Transport Is Modulated by p38 Kinase–Dependent Cross-Talk between ENaC and Na,K-ATPase in Collecting Duct Principal Cells
    Article Snippet: The equivalent short-circuit current across polar- ized cell monolayers was measured at the end of 48-hour Dox treatment and calculated according to Ohm’s law from the values of transepithelial potential and resistance measured with a Millicell device (Millipore), as previously described.15 Amiloride-sensitive and ouabain-sensitive short- circuit current were determined as the difference between values measured before and after incubation for 30 minutes with 1025 M benzamil added to the apical medium or 531025 M oua- bain added to the basal medium, respectively. .. Equal amounts of protein from lysed cultured cells or pools of 50microdissected ratCCDswere separated by 4%–12% SDS-PAGE (Invitrogen, Basel, Switzerland), and transferred to polyvi- nylidene difluoride membranes (Immobilon-P; Millipore, Bedford, MA), as previously described.36 Proteins of interest were detected by Western blotting using anti–g-ENaC antibody, diluted 1:104 (a gift of J. Löffing, University of Zürich, Switzerland); anti–glyceraldehyde 3- phosphate dehydrogenase (GAPDH) antibody, diluted 1:105 (Millipore); anti–E-cadherin antibody diluted 1:103 (BD Biosciences); anti–phospho p38 kinase (Thr180/Tyr182) and anti–p38 kinase antibodies diluted 1:103 (Cell Signaling Technology); Phospho-ATF-2 (Thr71) antibody diluted 1:103 (Cell Signaling Technology); anti– E-cadherin antibody diluted 1:103 (BD Transduction Laboratories); anti-phospho AMPK (Thr172) and anti-AMPK antibodies diluted 1:103 (Cell Signaling Technology); and anti–Na,K-ATPase a1-subunit antibody diluted 1:104.37 The antigen-antibody complexes were detected using Immobilon Western Chemiluminescent HRP Substrate (Millipore). ..

    Bioprocessing:

    Article Title: Adeno-associated virus vector mediated expression of an oncogenic retroviral envelope protein induces lung adenocarcinomas in immunocompetent mice.
    Article Snippet: .. Western Blot Analysis Monoclonal antibodies specific for phospho-EGFR (Tyr1068; Tyr1148), Akt, phospho-Akt (Thr308), phospho-Akt (Ser473), phospho-PDK1 (Ser241), phospho-GSK-3a/b (Ser21/9), phosphoCyclin D1 (Thr286), phospho-p44/42 MAPK (Thr202/Thr204), p44/42 MAPK, PTEN, phospho-PTEN (Ser380), phosphomTOR (Ser2448), phospho-p70S6K (Thr389), phospho-p38 MAPK (Thr180/Tyr182), phospho-ATF-2 (Thr71), phospho-STAT3 (Ser727), phospho-STAT3 (Tyr705), phospho-STAT5 (Tyr694), phospho-c-Fos (Ser32) and Cox2 were purchased from Cell Signaling Technology. .. Antibodies against surfactant protein C (SPC), Clara cell 10 (CC10) protein and actin were purchased from Santa Cruz Biotechnology.

    Cell Culture:

    Article Title: Sodium Transport Is Modulated by p38 Kinase–Dependent Cross-Talk between ENaC and Na,K-ATPase in Collecting Duct Principal Cells
    Article Snippet: The equivalent short-circuit current across polar- ized cell monolayers was measured at the end of 48-hour Dox treatment and calculated according to Ohm’s law from the values of transepithelial potential and resistance measured with a Millicell device (Millipore), as previously described.15 Amiloride-sensitive and ouabain-sensitive short- circuit current were determined as the difference between values measured before and after incubation for 30 minutes with 1025 M benzamil added to the apical medium or 531025 M oua- bain added to the basal medium, respectively. .. Equal amounts of protein from lysed cultured cells or pools of 50microdissected ratCCDswere separated by 4%–12% SDS-PAGE (Invitrogen, Basel, Switzerland), and transferred to polyvi- nylidene difluoride membranes (Immobilon-P; Millipore, Bedford, MA), as previously described.36 Proteins of interest were detected by Western blotting using anti–g-ENaC antibody, diluted 1:104 (a gift of J. Löffing, University of Zürich, Switzerland); anti–glyceraldehyde 3- phosphate dehydrogenase (GAPDH) antibody, diluted 1:105 (Millipore); anti–E-cadherin antibody diluted 1:103 (BD Biosciences); anti–phospho p38 kinase (Thr180/Tyr182) and anti–p38 kinase antibodies diluted 1:103 (Cell Signaling Technology); Phospho-ATF-2 (Thr71) antibody diluted 1:103 (Cell Signaling Technology); anti– E-cadherin antibody diluted 1:103 (BD Transduction Laboratories); anti-phospho AMPK (Thr172) and anti-AMPK antibodies diluted 1:103 (Cell Signaling Technology); and anti–Na,K-ATPase a1-subunit antibody diluted 1:104.37 The antigen-antibody complexes were detected using Immobilon Western Chemiluminescent HRP Substrate (Millipore). ..

    SDS Page:

    Article Title: Sodium Transport Is Modulated by p38 Kinase–Dependent Cross-Talk between ENaC and Na,K-ATPase in Collecting Duct Principal Cells
    Article Snippet: The equivalent short-circuit current across polar- ized cell monolayers was measured at the end of 48-hour Dox treatment and calculated according to Ohm’s law from the values of transepithelial potential and resistance measured with a Millicell device (Millipore), as previously described.15 Amiloride-sensitive and ouabain-sensitive short- circuit current were determined as the difference between values measured before and after incubation for 30 minutes with 1025 M benzamil added to the apical medium or 531025 M oua- bain added to the basal medium, respectively. .. Equal amounts of protein from lysed cultured cells or pools of 50microdissected ratCCDswere separated by 4%–12% SDS-PAGE (Invitrogen, Basel, Switzerland), and transferred to polyvi- nylidene difluoride membranes (Immobilon-P; Millipore, Bedford, MA), as previously described.36 Proteins of interest were detected by Western blotting using anti–g-ENaC antibody, diluted 1:104 (a gift of J. Löffing, University of Zürich, Switzerland); anti–glyceraldehyde 3- phosphate dehydrogenase (GAPDH) antibody, diluted 1:105 (Millipore); anti–E-cadherin antibody diluted 1:103 (BD Biosciences); anti–phospho p38 kinase (Thr180/Tyr182) and anti–p38 kinase antibodies diluted 1:103 (Cell Signaling Technology); Phospho-ATF-2 (Thr71) antibody diluted 1:103 (Cell Signaling Technology); anti– E-cadherin antibody diluted 1:103 (BD Transduction Laboratories); anti-phospho AMPK (Thr172) and anti-AMPK antibodies diluted 1:103 (Cell Signaling Technology); and anti–Na,K-ATPase a1-subunit antibody diluted 1:104.37 The antigen-antibody complexes were detected using Immobilon Western Chemiluminescent HRP Substrate (Millipore). ..



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    Image Search Results


    Effect of transient SUMOylation inhibition in pre-adipocytes on cAMP-PKA-p38 signaling and adaptive thermogenesis in mature adipocytes. ( A ) Heatmap showing the stable upregulation of most genes involved in adaptive thermogenesis and cAMP-PKA-p38 signaling 22 days after adipogenic induction. Z -scores were calculated and plotted in GraphPad Prism. Treatments of pre-adipocytes were performed as shown in Fig. . ( B ) Western blot analysis of PKA substrates phosphorylation, assessed using a pan-PKA-target antibody, 22 days after adipogenic induction. TBP was used as a loading control. The asterisk (*) indicates substrates significantly affected by TAK-981 and/or rosiglitazone treatment. ( C ) Quantification of PKA substrate signals from panel (B). The signal for PKA substrates and TBP was quantified using Fiji software, with PKA substrate signals normalized to TBP. Error bars represent the standard deviation of four independent experiments. Student’s t -test was used to calculate P -values. ( D ) Western blot analysis of p-CREB, p-ATF1, p38, p-p38, and p-ATF2, 22 days after adipogenic induction. TBP was used as a loading control. ( E ) Quantification of p-CREB/ATF-1 signals from panel (D). Signals were normalized to TBP. Error bars represent the standard deviation of three independent experiments. Student’s t -test was used to calculate P -values. ( F ) Western blot analysis of p-CREB in the absence or presence of the PKA inhibitor RP-8-CPT-cAMPS. TBP was used as a loading control. Quantification of p-CREB, normalized to TBP, is showed in the lower panel. Error bars represent the standard deviation of two independent experiments. Quantification of p-p38 ( G ) and p-ATF2 ( H ) from experiment in panel (D). Signals were normalized to TBP for p-ATF2 and to p38 for p-p38. Error bars represent the standard deviation of three independent experiments. Student’s t -test was used to calculate P -values.

    Journal: Nucleic Acids Research

    Article Title: Transient SUMOylation inhibition in human pre-adipocytes stably imprints a transcriptional beiging fate

    doi: 10.1093/nar/gkag232

    Figure Lengend Snippet: Effect of transient SUMOylation inhibition in pre-adipocytes on cAMP-PKA-p38 signaling and adaptive thermogenesis in mature adipocytes. ( A ) Heatmap showing the stable upregulation of most genes involved in adaptive thermogenesis and cAMP-PKA-p38 signaling 22 days after adipogenic induction. Z -scores were calculated and plotted in GraphPad Prism. Treatments of pre-adipocytes were performed as shown in Fig. . ( B ) Western blot analysis of PKA substrates phosphorylation, assessed using a pan-PKA-target antibody, 22 days after adipogenic induction. TBP was used as a loading control. The asterisk (*) indicates substrates significantly affected by TAK-981 and/or rosiglitazone treatment. ( C ) Quantification of PKA substrate signals from panel (B). The signal for PKA substrates and TBP was quantified using Fiji software, with PKA substrate signals normalized to TBP. Error bars represent the standard deviation of four independent experiments. Student’s t -test was used to calculate P -values. ( D ) Western blot analysis of p-CREB, p-ATF1, p38, p-p38, and p-ATF2, 22 days after adipogenic induction. TBP was used as a loading control. ( E ) Quantification of p-CREB/ATF-1 signals from panel (D). Signals were normalized to TBP. Error bars represent the standard deviation of three independent experiments. Student’s t -test was used to calculate P -values. ( F ) Western blot analysis of p-CREB in the absence or presence of the PKA inhibitor RP-8-CPT-cAMPS. TBP was used as a loading control. Quantification of p-CREB, normalized to TBP, is showed in the lower panel. Error bars represent the standard deviation of two independent experiments. Quantification of p-p38 ( G ) and p-ATF2 ( H ) from experiment in panel (D). Signals were normalized to TBP for p-ATF2 and to p38 for p-p38. Error bars represent the standard deviation of three independent experiments. Student’s t -test was used to calculate P -values.

    Article Snippet: The following antibodies were used: UCP1, abcam, ab209483; PKA phospho-substrates, Cell Signaling, 9624; p-CREB/p-ATF1, Cell Signaling, 9198; p38 MAPK, Cell Signaling, 9212; p-p38 MAPK, Cell Signaling, 9211; p-ATF2, Cell Signaling, 24329; SUMO2/3, abcam, ab81371; PPARG, Cell signaling, 2443; and TBP, Protein Tech, 22006-1-AP or abcam, 282715; γ-tubulin, Sigma, T5326: CEBPB, Santa Cruz, sc-7962 quantifications were performed using FiJi [ ].

    Techniques: Inhibition, Western Blot, Phospho-proteomics, Control, Software, Standard Deviation

    Journal: iScience

    Article Title: Negative regulation of lymphangiogenesis by Tenascin-C delays the resolution of inflammation

    doi: 10.1016/j.isci.2025.111756

    Figure Lengend Snippet:

    Article Snippet: Rabbit monoclonal anti-phospho ATF-2 (Thr71) (11G2) , CST , 5112; RRID: AB_560873.

    Techniques: Recombinant, Membrane, Blocking Assay, Western Blot, Ointment, cDNA Synthesis, Proliferation Assay, Software