Journal: JNCI Journal of the National Cancer Institute

Article Title: Immunotherapeutic Potential of Anti-Human Endogenous Retrovirus-K Envelope Protein Antibodies in Targeting Breast Tumors

doi: 10.1093/jnci/djr540

Figure Lengend Snippet: Effect of 6H5 monoclonal antibody (mAb) on apoptosis of breast cancer cells. A) Cells were treated with 6H5 mAb (red line) or control mIgG (gray line) (10 μg/mL of each antibody) for 16 hours, stained with annexin V–allophycocyanin and 7-AAD-phycoerythrin-cyanide 7, and analyzed by flow cytometry. B) Effect of 6H5 mAb treatment on cell death–inducing DFFA-like effector A (CIDEA) protein expression. Breast cancer cell lines were treated with 6H5 mAb or mIgG (10 μg/mL of each antibody) for 24 hours and analyzed for changes in protein expression by immunoblot using a mouse anti-human CIDEA antibody. ACTB was used as the protein loading control. Results are representative of two independent assays. C) The effect of 6H5 mAb treatment on expression of TP53 and TP53AIP1 proteins. MCF-7 and MDA-MB-231 breast cancer cell lines were treated with 6H5 mAb or mIgG (10 μg/mL of each antibody) for 24 hours, and an immunoblot assay was done using mouse anti-human TP53 and rabbit anti-human TP53AIP1 antibodies. ACTB was used as the protein loading control. Results are representative of two independent assays. D) Expression of active caspases 3 and 9 was assessed by immunoblot assay in ZR-75-1 and MDA-MB-231 breast cancer cells treated with 6H5 mAb or 6E11 mAb, or with mIgG (10 μg/mL of each antibody) for 24 hours using rabbit anti-human caspase 3 and mouse anti-human caspase 9 antibodies. ACTB was used as the protein loading control (top panel). Expression of active caspase 8 was assessed by immunoblot assay in MDA-MB-453 and MCF-7 breast cancer cells treated with 6H5 mAb (10 , 25, or 50 μg/mL), or with mIgG (10 μg/mL) for 24 hours using mouse anti-human caspase 8 antibody (bottom panel). Results are representative of at least two independent assays. E) Immunofluorescence assay to assess the expression of caspase proteins in MDA-MB-231 cells treated with 6H5 mAb or mIgG (10 μg/mL of each antibody) for 24 hours using rabbit anti-human caspase 3, mouse anti-human caspase 8, and mouse anti-human caspase 9 antibodies. Results are representative of two independent assays. Scale bar = 10 μm. F) Immunofluorescence assay to assess the expression of CDK5 and CDKN1A proteins. MDA-MB-231 cells were treated with 6H5 mAb or mIgG (10 μg/mL of each antibody) using mouse anti-human CDK5 and mouse anti-human CDKN1A antibodies. Results are representative of at least two independent assays. Scale bar = 10 μm.

Article Snippet: Mouse anti-human cell death–inducing DFFA-like effector A (CIDEA) antibody (Abnova, Taipei, Taiwan; 1:1000 dilution) was used for detection of CIDEA expression in cells treated with 6H5 mAb or mIgG.

Techniques: Control, Staining, Flow Cytometry, Expressing, Western Blot, Immunofluorescence

Journal: JNCI Journal of the National Cancer Institute

Article Title: Immunotherapeutic Potential of Anti-Human Endogenous Retrovirus-K Envelope Protein Antibodies in Targeting Breast Tumors

doi: 10.1093/jnci/djr540

Figure Lengend Snippet: Effect of anti-HERV-K antibody treatment on protein expression in breast cell lines, analyzed by flow cytometry *

Article Snippet: Mouse anti-human cell death–inducing DFFA-like effector A (CIDEA) antibody (Abnova, Taipei, Taiwan; 1:1000 dilution) was used for detection of CIDEA expression in cells treated with 6H5 mAb or mIgG.

Techniques: Expressing, Flow Cytometry

Journal: Journal of integrative medicine

Article Title: Alisol B 23-acetate promotes white adipose tissue browning to mitigate high-fat diet-induced obesity by regulating mTOR-SREBP1 signaling.

doi: 10.1016/j.joim.2024.01.003

Figure Lengend Snippet: Fig. 2. AB23A treatment increased white adipose tissue (WAT) browning markers in mice. A: H&E staining of iWAT (400). B and C: The expression fold change of WAT browning marker genes (B) and proteins (C) in iWAT. In B, n = 6. *P < 0.05, vs NCD group; #P < 0.05, vs HFD group. Data are presented as mean ± standard deviation. AB23A: alisol B 23-acetate; CIDEA: cell death inducing DNA fragmentation factor-a-like effector A; H&E: hematoxylin and eosin; HFD: high-fat diet; iWAT: inguinal white adipose tissue; NCD: normal chow diet; PGC-1a: peroxisome proliferator-activated receptor-c coactivator 1a; PRDM-16: positive regulatory domain-containing 16; UCP1: uncoupling protein 1.

Article Snippet: The membranes were incubated overnight at 4 °C with the corresponding primary antibodies, including peroxisome proliferator-activated receptor-c coactivator 1a (PGC1a), positive regulatory domain-containing 16 (PRDM-16), sterol regulatory element-binding protein-1 (SREBP1), mTOR (ab191838, ab303534, ab28481, and ab134903; Abcam, UK), UCP1, cell death inducing DNA fragmentation factor-a-like effector A (CIDEA) and b-actin (23673-1-AP, 13170-1-AP, and 81115–1-RR; Proteintech, China).

Techniques: Staining, Expressing, Marker, Standard Deviation

Journal: Journal of integrative medicine

Article Title: Alisol B 23-acetate promotes white adipose tissue browning to mitigate high-fat diet-induced obesity by regulating mTOR-SREBP1 signaling.

doi: 10.1016/j.joim.2024.01.003

Figure Lengend Snippet: Fig. 5. AB23A treatment promoted UCP1 expression in 3T3-L1 adipocytes. A: immunofluorescence images of UCP1 in 3T3-L1 cells; B and C: fold change of the expression of genes (B) and proteins (C) associated with white adipose tissue browning in 3T3-L1 cells. *P < 0.05, vs Con group; #P < 0.05, vs MDI group. Data are presented as mean ± standard deviation. AB23A: alisol B 23-acetate; CIDEA: cell death inducing DNA fragmentation factor-a-like effector A; Con: control; DAPI: 2-(4-amidinophenyl)-6- indolecarbamidine dihydrochloride; MDI: 3-isobutyl-1-methylxanthine, dexamethasone and insulin; PGC-1a: peroxisome proliferator-activated receptor-c coactivator 1a; PRDM-16: positive regulatory domain-containing 16; UCP1: uncoupling protein 1.

Article Snippet: The membranes were incubated overnight at 4 °C with the corresponding primary antibodies, including peroxisome proliferator-activated receptor-c coactivator 1a (PGC1a), positive regulatory domain-containing 16 (PRDM-16), sterol regulatory element-binding protein-1 (SREBP1), mTOR (ab191838, ab303534, ab28481, and ab134903; Abcam, UK), UCP1, cell death inducing DNA fragmentation factor-a-like effector A (CIDEA) and b-actin (23673-1-AP, 13170-1-AP, and 81115–1-RR; Proteintech, China).

Techniques: Expressing, Standard Deviation, Control

Journal: Journal of integrative medicine

Article Title: Alisol B 23-acetate promotes white adipose tissue browning to mitigate high-fat diet-induced obesity by regulating mTOR-SREBP1 signaling.

doi: 10.1016/j.joim.2024.01.003

Figure Lengend Snippet: Fig. 6. AB23A inhibited lipid accumulation by impeding the mTOR-SREBP1 signaling pathway. A: the signaling pathways influenced by AB23A; B–E: the expression of mTOR- SREBP1 pathway-related proteins; F and G: results of oil red O staining; H: protein expression of white adipose tissue browning markers in 3T3-L1 cells. *P < 0.05, vs Con group; #P < 0.05, vs MDI group; &P < 0.05, vs MDI + AB23A group. Data are presented as mean ± standard deviation. AB23A: alisol B 23-acetate; AMPK: adenosine monophosphate-activated protein kinase; CIDEA: cell death inducing DNA fragmentation factor-a-like effector A; Con: control; MDI: 3-isobutyl-1-methylxanthine, dexamethasone and insulin; mTOR: mammalian target of rapamycin; PGC-1a: peroxisome proliferator-activated receptor-c coactivator 1a; PRDM-16: positive regulatory domain-containing 16; SREBP1: sterol regulatory element-binding protein-1; UCP1: uncoupling protein 1.

Article Snippet: The membranes were incubated overnight at 4 °C with the corresponding primary antibodies, including peroxisome proliferator-activated receptor-c coactivator 1a (PGC1a), positive regulatory domain-containing 16 (PRDM-16), sterol regulatory element-binding protein-1 (SREBP1), mTOR (ab191838, ab303534, ab28481, and ab134903; Abcam, UK), UCP1, cell death inducing DNA fragmentation factor-a-like effector A (CIDEA) and b-actin (23673-1-AP, 13170-1-AP, and 81115–1-RR; Proteintech, China).

Techniques: Protein-Protein interactions, Expressing, Staining, Standard Deviation, Control, Binding Assay

Journal: Journal of integrative medicine

Article Title: Alisol B 23-acetate promotes white adipose tissue browning to mitigate high-fat diet-induced obesity by regulating mTOR-SREBP1 signaling.

doi: 10.1016/j.joim.2024.01.003

Figure Lengend Snippet: Fig. 2. AB23A treatment increased white adipose tissue (WAT) browning markers in mice. A: H&E staining of iWAT (400). B and C: The expression fold change of WAT browning marker genes (B) and proteins (C) in iWAT. In B, n = 6. *P < 0.05, vs NCD group; #P < 0.05, vs HFD group. Data are presented as mean ± standard deviation. AB23A: alisol B 23-acetate; CIDEA: cell death inducing DNA fragmentation factor-a-like effector A; H&E: hematoxylin and eosin; HFD: high-fat diet; iWAT: inguinal white adipose tissue; NCD: normal chow diet; PGC-1a: peroxisome proliferator-activated receptor-c coactivator 1a; PRDM-16: positive regulatory domain-containing 16; UCP1: uncoupling protein 1.

Article Snippet: The membranes were incubated overnight at 4 °C with the corresponding primary antibodies, including peroxisome proliferator-activated receptor-c coactivator 1a (PGC1a), positive regulatory domain-containing 16 (PRDM-16), sterol regulatory element-binding protein-1 (SREBP1), mTOR (ab191838, ab303534, ab28481, and ab134903; Abcam, UK), UCP1, cell death inducing DNA fragmentation factor-a-like effector A (CIDEA) and b-actin (23673-1-AP, 13170-1-AP, and 81115–1-RR; Proteintech, China).

Techniques: Staining, Expressing, Marker, Standard Deviation

Journal: Journal of integrative medicine

Article Title: Alisol B 23-acetate promotes white adipose tissue browning to mitigate high-fat diet-induced obesity by regulating mTOR-SREBP1 signaling.

doi: 10.1016/j.joim.2024.01.003

Figure Lengend Snippet: Fig. 5. AB23A treatment promoted UCP1 expression in 3T3-L1 adipocytes. A: immunofluorescence images of UCP1 in 3T3-L1 cells; B and C: fold change of the expression of genes (B) and proteins (C) associated with white adipose tissue browning in 3T3-L1 cells. *P < 0.05, vs Con group; #P < 0.05, vs MDI group. Data are presented as mean ± standard deviation. AB23A: alisol B 23-acetate; CIDEA: cell death inducing DNA fragmentation factor-a-like effector A; Con: control; DAPI: 2-(4-amidinophenyl)-6- indolecarbamidine dihydrochloride; MDI: 3-isobutyl-1-methylxanthine, dexamethasone and insulin; PGC-1a: peroxisome proliferator-activated receptor-c coactivator 1a; PRDM-16: positive regulatory domain-containing 16; UCP1: uncoupling protein 1.

Article Snippet: The membranes were incubated overnight at 4 °C with the corresponding primary antibodies, including peroxisome proliferator-activated receptor-c coactivator 1a (PGC1a), positive regulatory domain-containing 16 (PRDM-16), sterol regulatory element-binding protein-1 (SREBP1), mTOR (ab191838, ab303534, ab28481, and ab134903; Abcam, UK), UCP1, cell death inducing DNA fragmentation factor-a-like effector A (CIDEA) and b-actin (23673-1-AP, 13170-1-AP, and 81115–1-RR; Proteintech, China).

Techniques: Expressing, Standard Deviation, Control

Journal: Journal of integrative medicine

Article Title: Alisol B 23-acetate promotes white adipose tissue browning to mitigate high-fat diet-induced obesity by regulating mTOR-SREBP1 signaling.

doi: 10.1016/j.joim.2024.01.003

Figure Lengend Snippet: Fig. 6. AB23A inhibited lipid accumulation by impeding the mTOR-SREBP1 signaling pathway. A: the signaling pathways influenced by AB23A; B–E: the expression of mTOR- SREBP1 pathway-related proteins; F and G: results of oil red O staining; H: protein expression of white adipose tissue browning markers in 3T3-L1 cells. *P < 0.05, vs Con group; #P < 0.05, vs MDI group; &P < 0.05, vs MDI + AB23A group. Data are presented as mean ± standard deviation. AB23A: alisol B 23-acetate; AMPK: adenosine monophosphate-activated protein kinase; CIDEA: cell death inducing DNA fragmentation factor-a-like effector A; Con: control; MDI: 3-isobutyl-1-methylxanthine, dexamethasone and insulin; mTOR: mammalian target of rapamycin; PGC-1a: peroxisome proliferator-activated receptor-c coactivator 1a; PRDM-16: positive regulatory domain-containing 16; SREBP1: sterol regulatory element-binding protein-1; UCP1: uncoupling protein 1.

Article Snippet: The membranes were incubated overnight at 4 °C with the corresponding primary antibodies, including peroxisome proliferator-activated receptor-c coactivator 1a (PGC1a), positive regulatory domain-containing 16 (PRDM-16), sterol regulatory element-binding protein-1 (SREBP1), mTOR (ab191838, ab303534, ab28481, and ab134903; Abcam, UK), UCP1, cell death inducing DNA fragmentation factor-a-like effector A (CIDEA) and b-actin (23673-1-AP, 13170-1-AP, and 81115–1-RR; Proteintech, China).

Techniques: Protein-Protein interactions, Expressing, Staining, Standard Deviation, Control, Binding Assay