antibodies against phospho ampk t172  (Cell Signaling Technology Inc)

Journal: Redox report : communications in free radical research

Article Title: Rotenone inhibited osteosarcoma metastasis by modulating ZO-2 expression and location via the ROS/Ca 2+ /AMPK pathway.

doi: 10.1080/13510002.2025.2493556

Figure Lengend Snippet: Figure 1. Rotenone-induced ROS inhibited OS metastasis. (A) IC50 of Rotenone in MG-63, U2OS and 143B cells was measured by MTT assay (0.02, 1.09 and 1.35 μM, respectively). (B) Intracellular ROS levels after OS cells were treated with Rotenone (0.02 and 1.09 μM Rotenone for MG-63 and U2OS respectively for 48 h) or Rotenone + NAC (10 mM). (C–E) Metastasis inhibition effects of Rotenone were measured by wound-healing and transwell assays. (F) Rotenone modulated OS cell AMPK activity, ZO-2 expression, and EMT, as determined via Western blot analysis of EMT-related markers. (G) Bar graph depicting quantitative analysis of Western blots. *p < 0.05, **p < 0.01, ***p < 0.001 vs. control.

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against p-AMPK-T172 (2535, CST, USA) and ZO-2 (ab224314, Abcam, USA).

Techniques: MTT Assay, Inhibition, Activity Assay, Expressing, Western Blot, Control

Journal: Redox report : communications in free radical research

Article Title: Rotenone inhibited osteosarcoma metastasis by modulating ZO-2 expression and location via the ROS/Ca 2+ /AMPK pathway.

doi: 10.1080/13510002.2025.2493556

Figure Lengend Snippet: Figure 2. Rotenone inhibited Ca2+/AMPK/ZO-2 signaling. (A) Intracellular Ca2+ levels after OS cells were treated with Rotenone (0.02 and 1.09 μM Rotenone for MG-63 and U2OS respectively for 48 h) or Rotenone + NAC (10 mM). (B–D) Metastasis capacity of different groups was measured by wound-healing and transwell assays. (E) AMPK activity, ZO-2 expression, and EMT-related markers were determined by Western blot analysis of OS cells treated with the indicated conditions. (F) Bar graph depicting quantitative analysis of Western blots. ***p < 0.001 vs. control.

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against p-AMPK-T172 (2535, CST, USA) and ZO-2 (ab224314, Abcam, USA).

Techniques: Activity Assay, Expressing, Western Blot, Control

Journal: Redox report : communications in free radical research

Article Title: Rotenone inhibited osteosarcoma metastasis by modulating ZO-2 expression and location via the ROS/Ca 2+ /AMPK pathway.

doi: 10.1080/13510002.2025.2493556

Figure Lengend Snippet: Figure 3. Rotenone and AMPK modulated ZO-2 subcellular location. (A, B) Immunoblotting and IF analysis of the subcellular distribution of ZO-2 after Rotenone treatment (0.02 and 1.09 μM Rotenone for MG-63 and U2OS respectively for 48 h). (C, D) AMPK inhibitor Compound C (10 μM for 48 h) modulated ZO-2 nuclear translocation, as measured by immunoblotting and IF. (E) Co-IP was employed to reveal the interaction between AMPK and ZO-2.

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against p-AMPK-T172 (2535, CST, USA) and ZO-2 (ab224314, Abcam, USA).

Techniques: Western Blot, Translocation Assay, Co-Immunoprecipitation Assay

Journal: Redox report : communications in free radical research

Article Title: Rotenone inhibited osteosarcoma metastasis by modulating ZO-2 expression and location via the ROS/Ca 2+ /AMPK pathway.

doi: 10.1080/13510002.2025.2493556

Figure Lengend Snippet: Figure 4. Rotenone inhibited OS progression in vivo. (A, B) Rotenone suppressed tumor growth over time and showed a synergistic effect with Doxorubicin. Tumors were measured on the indicated days using Vernier calipers. Body weights of tumor-bearing mice were measured. (C, D) The expression of p-AMPK and ZO-2 in xenograft tumor tissues was determined by immunoblotting and IHC. (A, E) Rotenone suppressed lung metastasis and exhibited higher efficacy when combined with Doxorubicin. n = 5. The black dots on the surface of the lungs indicate metastasis foci. *p < 0.05, ***p < 0.001 vs. control. Rotenone and Doxorubicin administration was described in Materials and Methods.

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against p-AMPK-T172 (2535, CST, USA) and ZO-2 (ab224314, Abcam, USA).

Techniques: In Vivo, Expressing, Western Blot, Control

Journal: Redox report : communications in free radical research

Article Title: Rotenone inhibited osteosarcoma metastasis by modulating ZO-2 expression and location via the ROS/Ca 2+ /AMPK pathway.

doi: 10.1080/13510002.2025.2493556

Figure Lengend Snippet: Figure 5. ZO-2 expression was correlated with p-AMPK and metastasis in OS patients. (A) Expression of p-AMPK and ZO-2 was measured by IHC in metastatic and non-metastatic OS specimens. (B) The GEPIA database showed a positive correlation between p-AMPK and ZO-2 in sarcoma samples. (C) Working model illustrat ing Rotenone modulation of ZO-2 expression and location via the ROS/Ca2+/AMPK pathway.

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against p-AMPK-T172 (2535, CST, USA) and ZO-2 (ab224314, Abcam, USA).

Techniques: Expressing

Journal: International journal of molecular sciences

Article Title: Astaxanthin Alleviates Oxidative Stress in Mouse Preantral Follicles and Enhances Follicular Development Through the AMPK Signaling Pathway.

doi: 10.3390/ijms26052241

Figure Lengend Snippet: Figure 4. Effects of astaxanthin on expression of AMPK pathway proteins, mitochondrial biogenesis proteins, and antioxidant proteins. (A) Western blot analysis of expression of AMPK pathway proteins (p-AMPK and PGC-1α), mitochondrial biogenesis proteins (NRF1 and TFAM), and antioxidant proteins (NRF2 and HO-1) in follicles on day 10. (B) Analysis of relative expression levels of these proteins using Image J image analysis software. Experiment was repeated three times independently (N = 3), and 40 follicles were used in each group. * p < 0.05, compared with control group. # p < 0.05, compared with astaxanthin group.

Article Snippet: 2025, 26, 2241 16 of 21 the membrane was blocked with 5% non-fat milk for 1 h. After blocking, the membrane was incubated overnight at 4 ◦C with primary antibodies against p-AMPK (#50081, Cell Signaling Technology, Boston, MA, USA), PGC-1α (#ab191838, Abcam, Cambridge, UK), NRF1 (#46743, Cell Signaling Technology), TFAM (#ab252432, Abcam), NRF2 (#ab92946, Abcam), HO-1 (#ab52947, Abcam), PINK1 (#ab23707, Abcam), Parkin (#32833, Cell Signaling Technology), LC3-II (#43566, Cell Signaling Technology), cleaved caspase 3 (#9664, Cell Signaling Technology), Bax (#2772, Cell Signaling Technology), Bcl-2 (#3498, Cell Signaling Technology), P53 (#ab26, Abcam), StAR (#ab203193, Abcam), P450scc (#ab272494, Abcam), and β-actin (#ab8226, Abcam).

Techniques: Expressing, Western Blot, Software, Control