Journal: Cell Death & Disease

Article Title: The DAG/PKC/CREB1/TGF-β1 axis drives shear-wave elastography stiffness and malignant progression in triple-negative breast cancer via lipid metabolic reprogramming

doi: 10.1038/s41419-026-08625-0

Figure Lengend Snippet: A Cell viability measured by CCK-8 assay in MDA-MB-231 and E0771 cells treated with increasing concentrations of Sotrastaurin. Western blot analysis of PKC, phosphorylated PKC (p-PKC), and epithelial–mesenchymal transition (EMT)-related markers in MDA-MB-231 ( B ) and E0771 ( C ) cells after treatment with Diolein and Sotrastaurin. D BODIPY staining and quantitative analysis of lipid droplets in MDA-MB-231 cells treated with Diolein. Scale bars, 50 μm. Quantitative results of colony formation assays in MDA-MB-231 ( E ) and E0771 ( F ) cells treated with Diolein and Sotrastaurin. Wound healing assay and quantitative analysis of migration in MDA-MB-231 ( G ) and E0771 ( H ) cells treated with Diolein and Sotrastaurin. Scale bars, 100 μm. Transwell invasion and migration assays with quantitative results in MDA-MB-231 ( I ) and E0771 ( J ) cells after treatment with Diolein and Sotrastaurin. Scale bars, 100 μm. Data are presented as mean ± SEM.

Article Snippet: Human BC cell line MDA-MB-231 and murine BC cell lines E0771, 4T1, and AT3 were purchased from the American Type Culture Collection (ATCC).

Techniques: CCK-8 Assay, Western Blot, Staining, Wound Healing Assay, Migration

Journal: Cell Death & Disease

Article Title: The DAG/PKC/CREB1/TGF-β1 axis drives shear-wave elastography stiffness and malignant progression in triple-negative breast cancer via lipid metabolic reprogramming

doi: 10.1038/s41419-026-08625-0

Figure Lengend Snippet: A CREB1 mRNA expression was downregulated in Sotrastaurin-treated MDA-MB-231 cells compared to controls, as determined by RNA-seq. B Analysis of the GEO dataset GSE137467 confirmed that PKC inhibitor treatment also reduced CREB1 mRNA levels in this cell line. Western blot analysis of CREB1 and phosphorylated CREB1 (p-CREB1) in MDA-MB-231 ( C ) and E0771 ( D ) cells treated with Diolein and Sotrastaurin. E TGF-β1 mRNA expression was downregulated in Sotrastaurin-treated MDA-MB-231 cells compared to controls, as determined by RNA-seq. F Western blot analysis of CREB1, p-CREB1, TGF-β1, MMP9 and N-cadherin in MDA-MB-231 treated with siCREB1 and Diolein. G Western blot analysis of CREB1, p-CREB1, and TGF-β1 in MDA-MB-231 ( G ) and E0771 ( H ) cells treated with PMA (PKC activator) and 666-15 (CREB1 inhibitor). I Western blot analysis of EMT-related markers in MDA-MB-231 cells treated with 666-15 and SRI-011381 (TGF-β1 activator). Quantitative results of colony formation assays ( J ), wound healing assay ( K ) and transwell invasion and migration assays ( L ) in MDA-MB-231 cells treated with siCREB1 and Diolein. Scale bars 100 μm. Quantitative results of colony formation assays in MDA-MB-231 ( M ) and E0771 ( N ) cells treated with 666-15 and SRI-011381. Wound healing assay and quantitative analysis of migration in MDA-MB-231 ( O ) and E0771 ( P ) cells treated with 666-15 and SRI-011381. Scale bars, 100 μm. Transwell invasion and migration assays with quantitative results in MDA-MB-231 ( Q ) and E0771 ( R ) cells after treatment with 666-15 and SRI-011381. Scale bars, 100μm. Data are presented as mean ± SEM.

Article Snippet: Human BC cell line MDA-MB-231 and murine BC cell lines E0771, 4T1, and AT3 were purchased from the American Type Culture Collection (ATCC).

Techniques: Expressing, RNA Sequencing, Western Blot, Wound Healing Assay, Migration

Journal: Cell Death & Disease

Article Title: The DAG/PKC/CREB1/TGF-β1 axis drives shear-wave elastography stiffness and malignant progression in triple-negative breast cancer via lipid metabolic reprogramming

doi: 10.1038/s41419-026-08625-0

Figure Lengend Snippet: A Western blot analysis of COL1A1 and α-SMA expression in NIH-3T3 cells treated with control, conditioned medium from Diolein-treated E0771 cells (CM(D-7)), or CM(D-7) supplemented with P144 (TGF-β1 inhibitor). B Immunofluorescence staining and quantitative analysis of COL1A1 and α-SMA in NIH-3T3 cells under the same treatment conditions as in ( A ). Scale bar, 50μm. C TGF-β1 secretion levels measured by ELISA in NIH-3T3 cells cultured with different conditioned media. D Phalloidin staining of E0771 cells treated with control, conditioned medium from NIH-3T3 exposed to CM(D-7) (CM(D-7-3)), CM(D-7-3) + P144, or recombinant TGF-β1. Scale bar, 20μm. E BODIPY staining and quantitative analysis of lipid droplets in E0771 cells treated with control, Diolein, conditioned medium from NIH-3T3 (CM(3)), or CM(D-7-3). Scale bar, 50 μm. F Measurement of triglyceride and cholesterol levels in E0771 cells treated with control or CM(D-7-3). G Western blot analysis of lipid metabolism markers (FASN, SREBP, PPARγ) in E0771 cells treated with control, CM(3), conditioned medium from E0771-exposed NIH-3T3 (CM(7-3)), or CM(D-7-3). H Transwell invasion and migration assays with quantitative results in MDA-MB-231 and E0771 cells treated with control or CM(D-7-3). Scale bar, 100 μm. Data are presented as mean ± SEM.

Article Snippet: Human BC cell line MDA-MB-231 and murine BC cell lines E0771, 4T1, and AT3 were purchased from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Expressing, Control, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Cell Culture, Recombinant, Migration

Journal: Cell Death & Disease

Article Title: The DAG/PKC/CREB1/TGF-β1 axis drives shear-wave elastography stiffness and malignant progression in triple-negative breast cancer via lipid metabolic reprogramming

doi: 10.1038/s41419-026-08625-0

Figure Lengend Snippet: A SWE and B-mode ultrasound images of tumors from the E0771 model. ( n = 10 per group). Quantitative analysis of SWE-based tumor stiffness ( B ), LOX levels (measured by ELISA) ( C ), α-SMA IHC ( D ) and Sirius Red staining ( E ) in tumor tissues from the E0771 model. Quantitative analysis of SWE-based tumor stiffness ( F ), LOX levels (measured by ELISA) ( G ), α-SMA IHC ( H ) and Sirius Red staining ( I ) in tumor tissues from the AT3 model. Quantitative analysis of SWE-based tumor stiffness ( J ), LOX levels (measured by ELISA) ( K ), α-SMA IHC ( L ) and Sirius Red staining ( M ) in tumor tissues from the MDA-MB-231 model. Data are presented as mean ± SEM.

Article Snippet: Human BC cell line MDA-MB-231 and murine BC cell lines E0771, 4T1, and AT3 were purchased from the American Type Culture Collection (ATCC).

Techniques: Enzyme-linked Immunosorbent Assay, Staining