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. 2023 Dec 11;14(12):815. doi: 10.1038/s41419-023-06352-4

Fig. 7. Mint3 depletion induces energy stress and thereby attenuates HSF-1 activity in TNBC tumors.

Fig. 7

A, B Immunostaining of hypoxic markers pimonidazole (green) and endothelial cell marker CD31 (red) in MDA-MB-231 (A) and MDA-MB-468 (B) ishMint3 tumors administered with or without doxycycline (DOX) for 3 days. Nuclei were counterstained with Hoechst 33342 (blue). C, D Lactate levels in MDA-MB-231 (C) and MDA-MB-468 (D) ishMint3 tumors administered with or without DOX for three days. n = 10 per group. E, F ATP levels in MDA-MB-231 (E) and MDA-MB-468 (F) ishMint3 tumors administered with or without DOX for 3 days. n = 10 per group. G, H Phosphorylation levels of AMPK1α in MDA-MB-231 (G) and MDA-MB-468 (H) ishMint3 tumors administered with or without DOX for 3 days. n = 3 per group. I Immunostaining of hypoxic marker pimonidazole (green) and phospho-AMPK1α or phospho-HSF-1 Ser326 (red) in MDA-MB-231 and MDA-MB-468 tumors. Nuclei were counterstained with Hoechst 33342 (blue). J Immunostaining of hypoxic marker pimonidazole (green) and endothelial cell marker CD31 (red) in MDA-MB-231 tumors administered with or without a glycolysis inhibitor 2-deoxyglucose (2-DG; 500 mg/kg b.w.) for 3 days. Nuclei were counterstained with Hoechst 33342 (blue). K ATP levels in MDA-MB-231 tumors were administered with or without 2-DG for 3 days. n = 6 per group. L Immunoblotting of Mint3, HSF-1, HSP70, and mTOR signaling-related proteins in MDA-MB-231 tumors administered with or without 2-DG for 3 days. n = 3 per group. In C–F and K, the data represent the mean ± SEM and were analyzed using the Mann–Whitney U-test. **p < 0.01, ***p < 0.001, ****p < 0.0001. NS not significant.