p atf2  (Cell Signaling Technology Inc)

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: bioRxiv

Article Title: Clove Aqueous Extract Triggers a Multi-Organellar Stress Crisis through Lysosomal Destabilisation and Mitochondrial Hyperpolarisation to Suppress Patient-Derived Ovarian Cancer Cells

doi: 10.64898/2026.01.28.702206

Figure Lengend Snippet: Quantification of P-ATF-2 level in (A) OCAS12, (B) OCAS14, (C) OCAS17, and (D) OCAST16 cells following treatment with CAE. Data are normalised to the non-treated (NT) control and presented as mean ± SEM of n=3 independent experiments. Statistical significance was determined by one-way ANOVA with a post-hoc Tukey test. *p < 0.05, **p < 0.01, ***p < 0.001, ns = not significant.

Article Snippet: Exposed cells were then washed in PBS, fixed with 3% PFA and stained for either ATF-2 and/or p-ATF-2 (Santa Cruz) and nuclei (Hoechst).

Techniques: Control