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p creb p atf1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p creb p atf1
    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, <t>p-ATF1,</t> 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.
    P Creb P Atf1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 2146 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p+creb+p+atf1/Phospho-CREB+(Ser133)+Rabbit+mAb/pmc13000455-190-13-14
    Average 97 stars, based on 2146 article reviews
    p creb p atf1 - by Bioz Stars, 2026-09
    97/100 stars

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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

    Related Articles

    Inhibition:

    Article Title: Transient SUMOylation inhibition in human pre-adipocytes stably imprints a transcriptional beiging fate
    Article Snippet: Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.. 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 [ ].

    Article Title: SUMOylation Restricts Adaptive Thermogenesis by Suppressing Beiging Gene Networks
    Article Snippet: 15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.. The following antibodies were used: UCP1, abcam, ab209483; PKA phosphor-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 and TBP, Protein Tech, 22006-1-AP).. Quantifications were performed using FiJi ( Schindelin et al , 2012 ).Quantifications were performed using FiJi ( Schindelin et al , 2012 ).

    Western Blot:

    Article Title: Transient SUMOylation inhibition in human pre-adipocytes stably imprints a transcriptional beiging fate
    Article Snippet: Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.. 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 [ ].

    Article Title: SUMOylation Restricts Adaptive Thermogenesis by Suppressing Beiging Gene Networks
    Article Snippet: 15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.. The following antibodies were used: UCP1, abcam, ab209483; PKA phosphor-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 and TBP, Protein Tech, 22006-1-AP).. Quantifications were performed using FiJi ( Schindelin et al , 2012 ).Quantifications were performed using FiJi ( Schindelin et al , 2012 ).

    Phospho-proteomics:

    Article Title: Transient SUMOylation inhibition in human pre-adipocytes stably imprints a transcriptional beiging fate
    Article Snippet: Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.. 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 [ ].

    Article Title: SUMOylation Restricts Adaptive Thermogenesis by Suppressing Beiging Gene Networks
    Article Snippet: 15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.. The following antibodies were used: UCP1, abcam, ab209483; PKA phosphor-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 and TBP, Protein Tech, 22006-1-AP).. Quantifications were performed using FiJi ( Schindelin et al , 2012 ).Quantifications were performed using FiJi ( Schindelin et al , 2012 ).

    Control:

    Article Title: Transient SUMOylation inhibition in human pre-adipocytes stably imprints a transcriptional beiging fate
    Article Snippet: Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.. 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 [ ].

    Article Title: SUMOylation Restricts Adaptive Thermogenesis by Suppressing Beiging Gene Networks
    Article Snippet: 15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.. The following antibodies were used: UCP1, abcam, ab209483; PKA phosphor-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 and TBP, Protein Tech, 22006-1-AP).. Quantifications were performed using FiJi ( Schindelin et al , 2012 ).Quantifications were performed using FiJi ( Schindelin et al , 2012 ).

    Software:

    Article Title: Transient SUMOylation inhibition in human pre-adipocytes stably imprints a transcriptional beiging fate
    Article Snippet: Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.. 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 [ ].

    Article Title: SUMOylation Restricts Adaptive Thermogenesis by Suppressing Beiging Gene Networks
    Article Snippet: 15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.. The following antibodies were used: UCP1, abcam, ab209483; PKA phosphor-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 and TBP, Protein Tech, 22006-1-AP).. Quantifications were performed using FiJi ( Schindelin et al , 2012 ).Quantifications were performed using FiJi ( Schindelin et al , 2012 ).

    Standard Deviation:

    Article Title: Transient SUMOylation inhibition in human pre-adipocytes stably imprints a transcriptional beiging fate
    Article Snippet: Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.Protein concentrations were determined with the Bradford assay and 30 μg protein were used for SDS–PAGE and western blot.. 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 [ ].

    Article Title: SUMOylation Restricts Adaptive Thermogenesis by Suppressing Beiging Gene Networks
    Article Snippet: 15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.15 – 50µg of protein were used for SDS-PAGE and proteins of interested detected by Western Blots.. The following antibodies were used: UCP1, abcam, ab209483; PKA phosphor-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 and TBP, Protein Tech, 22006-1-AP).. Quantifications were performed using FiJi ( Schindelin et al , 2012 ).Quantifications were performed using FiJi ( Schindelin et al , 2012 ).



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    Cell Signaling Technology Inc p creb p atf1
    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, <t>p-ATF1,</t> 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.
    P Creb P Atf1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p+creb+p+atf1/Phospho-CREB+(Ser133)+Rabbit+mAb/pmc13000455-190-13-14
    Average 97 stars, based on 1 article reviews
    p creb p atf1 - by Bioz Stars, 2026-09
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    p creb atf1  (Cell Signaling Technology Inc)
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    Cell Signaling Technology Inc p creb atf1
    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, <t>p-ATF1,</t> 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.
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    Average 86 stars, based on 1 article reviews
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    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