Journal: Scientific Reports

Article Title: Bcl-x L inhibition enhances Dinaciclib-induced cell death in soft-tissue sarcomas

doi: 10.1038/s41598-019-40106-7

Figure Lengend Snippet: List of primary antibodies employed for western blotting and/or immunofluorescence.

Article Snippet: p16 INK4a (CDKN2A) , Abcam , ab108349 , 1:1000.

Techniques: Western Blot, Immunofluorescence, Transduction

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: IL-17A promotes fatty acid uptake through the IL-17A/IL-17RA/p-STAT3/FABP4 axis to fuel ovarian cancer growth in an adipocyte-rich microenvironment

doi: 10.1007/s00262-019-02445-2

Figure Lengend Snippet: IL-17A increased the OvCa growth and metastasis in the peritoneal cavity of a murine model. ID8 cells (murine OvCa) were prepared for orthotopic/intrabursal injection into C57BL/6 WT and IL-17A−/− mice. Eight weeks after injection, the mice were killed. a, c Representative photos of ovarian tissues in the ID8-injected side (indicated by black arrow). b, d Representative photos of tumor nodules distributed in the abdominal cavity. a Omentum. b Bowel. c Mesentery. d Abdominal wall. e Number of tumor nodules in WT and IL-17A−/− mice. **p < 0.01. f Protein lysates were prepared from the tumor tissues of WT and IL-17A−/− mice, and the protein levels of FABP4, p-STAT3 and STAT3 were analyzed by Western blotting. Three independent experiments were performed and a representative image is shown. Data represent mean ± SD from three independent experiments. **p < 0.01. g The sections were prepared from the tumor tissues of WT and IL-17A−/− mice, and IHC staining was performed to determine FABP4 expression was performed. Three independent experiments were performed, and the representative image is shown

Article Snippet: Equal amounts of proteins were separated by 10–13% SDS-PAGE, transferred onto PVDF membrane and incubated at 4 °C overnight with specific primary antibodies against CD36 (Cat# 18836-1-AP) from Proteintech and FABP4 (Cat# DF6035), STAT3 (Cat# AF6294), p-STAT3 (Tyr705,Cat# AF3293) and GAPDH (Cat# AF7021) from Affinity.

Techniques: Injection, Western Blot, Immunohistochemistry, Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: IL-17A promotes fatty acid uptake through the IL-17A/IL-17RA/p-STAT3/FABP4 axis to fuel ovarian cancer growth in an adipocyte-rich microenvironment

doi: 10.1007/s00262-019-02445-2

Figure Lengend Snippet: rhIL-17A enhanced the proliferation of OvCa cells in the presence of PA via the IL-17A/IL-17RA/p-STAT3/FABP4 axis. a Proliferation assays of 2780 and OVCAR3 cells after treatment with rhIL-17A, PA, or rhIL-17A and PA for designated time periods. Cell proliferation was detected by MTS assay. b Proliferation assay for 2780 and OVCAR3 cells after pretreatment with BMS309403 or STATTIC for 2 h and then treatment with rhIL-17A, PA, or rhIL-17A and PA for 48 h. Data represent the mean ± SD from three independent experiments. *p < 0.05, **p < 0.01

Article Snippet: Equal amounts of proteins were separated by 10–13% SDS-PAGE, transferred onto PVDF membrane and incubated at 4 °C overnight with specific primary antibodies against CD36 (Cat# 18836-1-AP) from Proteintech and FABP4 (Cat# DF6035), STAT3 (Cat# AF6294), p-STAT3 (Tyr705,Cat# AF3293) and GAPDH (Cat# AF7021) from Affinity.

Techniques: MTS Assay, Proliferation Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: IL-17A promotes fatty acid uptake through the IL-17A/IL-17RA/p-STAT3/FABP4 axis to fuel ovarian cancer growth in an adipocyte-rich microenvironment

doi: 10.1007/s00262-019-02445-2

Figure Lengend Snippet: rhIL-17A increased the uptake of PA in OvCa cells via the IL-17A/IL-17RA/p-STAT3/FABP4 axis. A2780 and OVCAR3 cells were treated with rhIL-17A, PA, rhIL-17A and PA, BMS309403 or STATTIC in parental cells or control/IL-17RA-siRNA cells. After treatments, oil-red O staining was performed. a Bar graph for OD490 values (indicating the PA level inside the cells) from each group. Data represent the mean ± SD from three independent experiments. *p < 0.05, **p < 0.01. b Representative photos of oil-red O staining for each group. Magnification, ×100. 1: local enlarged image of A2780 cells after treatment with rhIL-17A and PA (shown in inset); 2: local enlarged image of OVCAR3 cells after treatment with rhIL-17A and PA (shown in inset). c Bodipy-FL-C16 capture assay after pretreatment with BMS309403 for 2 h and then rhIL-17A for 6 h in A2780, OVCAR3 and SKOV3 cells. Three independent experiments were performed and a representative image is shown

Article Snippet: Equal amounts of proteins were separated by 10–13% SDS-PAGE, transferred onto PVDF membrane and incubated at 4 °C overnight with specific primary antibodies against CD36 (Cat# 18836-1-AP) from Proteintech and FABP4 (Cat# DF6035), STAT3 (Cat# AF6294), p-STAT3 (Tyr705,Cat# AF3293) and GAPDH (Cat# AF7021) from Affinity.

Techniques: Control, Staining

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: IL-17A promotes fatty acid uptake through the IL-17A/IL-17RA/p-STAT3/FABP4 axis to fuel ovarian cancer growth in an adipocyte-rich microenvironment

doi: 10.1007/s00262-019-02445-2

Figure Lengend Snippet: Relationship between the levels of IL-17A and FABP4 in clinical OvCa settings. a Expression of IL-17A and FABP4 in normal ovary tissues and OvCa specimens, as determined by IHC staining and FIGO staging. Magnification, ×400; scale bar, 25 μm. Blue box: Typical characteristics of IL-17A+ve cells in an OvCa environment. 1. An IL-17A+ve small round lymphocyte characterized by a small round nucleus (enlarged in inset); 2. An IL-17A+ve large irregularly shaped macrophage with a kidney-shaped nucleus (shown in inset). b Percentage of FABP4 positivity in the IL-17A-negative/low (−/+) group and the IL-17A-high/very high (++/+++) group. The black bar represents the median of each group. A minimum of 10 fields per section was counted and analyzed. *p < 0.05

Article Snippet: Equal amounts of proteins were separated by 10–13% SDS-PAGE, transferred onto PVDF membrane and incubated at 4 °C overnight with specific primary antibodies against CD36 (Cat# 18836-1-AP) from Proteintech and FABP4 (Cat# DF6035), STAT3 (Cat# AF6294), p-STAT3 (Tyr705,Cat# AF3293) and GAPDH (Cat# AF7021) from Affinity.

Techniques: Expressing, Immunohistochemistry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: IL-17A promotes fatty acid uptake through the IL-17A/IL-17RA/p-STAT3/FABP4 axis to fuel ovarian cancer growth in an adipocyte-rich microenvironment

doi: 10.1007/s00262-019-02445-2

Figure Lengend Snippet: rhIL-17A increased FABP4 expression in OvCa cells via STAT3 signaling. Dose–effect (a, b) and time–effect (c) experiments were performed in A2780 and OVCAR3 cells. a mRNA level of FABP4 after rhIL-17A treatment. b, c Protein expression of FABP4 after rhIL-17A treatment. d-(a) Protein expression of FABP4, p-STAT3 and STAT3 after rhIL-17A and/or STATTIC treatment (A2780: 0.3125 μM; OVCAR3: 1.25 μM). d-(b) The relative expression of proteins in d-(a). Three independent experiments were performed and a representative image is shown. Data represent the mean ± SD from three independent experiments. *p < 0.05, **p < 0.01

Article Snippet: Equal amounts of proteins were separated by 10–13% SDS-PAGE, transferred onto PVDF membrane and incubated at 4 °C overnight with specific primary antibodies against CD36 (Cat# 18836-1-AP) from Proteintech and FABP4 (Cat# DF6035), STAT3 (Cat# AF6294), p-STAT3 (Tyr705,Cat# AF3293) and GAPDH (Cat# AF7021) from Affinity.

Techniques: Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: IL-17A promotes fatty acid uptake through the IL-17A/IL-17RA/p-STAT3/FABP4 axis to fuel ovarian cancer growth in an adipocyte-rich microenvironment

doi: 10.1007/s00262-019-02445-2

Figure Lengend Snippet: Proposed model for the mechanism by which IL-17A links adipocytes with OvCa cells in the ARM. In the ARM, IL-17A-producing cells secrete IL-17A, which upregulates FABP4 expression via p-STAT3 signaling. Meanwhile, adipocytes provide FAs, which are transported by FABP4 and are then utilized for ATP production by β-oxidization; subsequently, OvCa cell proliferation will be increased

Article Snippet: Equal amounts of proteins were separated by 10–13% SDS-PAGE, transferred onto PVDF membrane and incubated at 4 °C overnight with specific primary antibodies against CD36 (Cat# 18836-1-AP) from Proteintech and FABP4 (Cat# DF6035), STAT3 (Cat# AF6294), p-STAT3 (Tyr705,Cat# AF3293) and GAPDH (Cat# AF7021) from Affinity.

Techniques: Expressing

Journal: Biomolecules & Therapeutics

Article Title: Taurine Synthesis by 2-Aminoethanethiol Dioxygenase as a Vulnerable Metabolic Alteration in Pancreatic Cancer

doi: 10.4062/biomolther.2024.086

Figure Lengend Snippet: Inhibition of ADO mitigates pancreatic cancer cell (PANC-1) growth in vitro . (A) Relative expression of ADO in PANC-1 cells after control, ADO-KD1, and ADO-KD2 siRNA transfection (n=3). Two-sided Welch’s t-test was used. (B) Relative level of taurine in PANC-1 cells after control, ADO-KD1, and ADO-KD2 transfection (n=5). Two-sided Welch’s t-test was used. A.U.=arbitrary unit. (C) (Left) Cell proliferation of PANC-1 after control, ADO-KD1, and ADO-KD2 siRNA transfection was detected by CCK assay (n=8). (Right) Relative cell proliferation to day 0. Two-sided Welch’s t-test was used. O.D.=optical density. (D) Clonogenic assay of PANC-1 cells after control and ADO-KD1 siRNA transfection (20-day cultivation, n=3). (E) Expression of NF-κB (p65 and phosphorylated p65) and MEK (MEK1 and phosphorylated MEK1/2) proteins in PANC-1 cells after control and ADO-KD1 siRNA transfection, as detected by western blot assay. Data are presented as the mean ± SD. * p <0.05, ** p <0.01 and *** p <0.001.

Article Snippet: Western blot experiment was conducted using primary antibodies against ADO (16479-1-AP, Proteintech, Rosemont, IL, USA), β-actin (sc-47778, Santa Cruz), p65 (sc-8008, Santa Cruz), phospho-p65 (#3033, CST, Danvers, MA, USA), MEK1 (#9124, CST), phospho-MEK1 (Thr286) (#9127, CST), phospho-MEK1/2 (Ser221) (#2338, CST), ERK1/2 (#9102, CST), and phospho-ERK1/2 (#4377, CST).

Techniques: Inhibition, In Vitro, Expressing, Control, Transfection, Clonogenic Assay, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Aspirin inhibits adipogenesis of tendon stem cells and lipids accumulation in rat injury tendon through regulating PTEN/PI3K/AKT signalling

doi: 10.1111/jcmm.14622

Figure Lengend Snippet: Effects of aspirin on the adipogenic differentiation of TSCs. A, TSCs were treated with increasing concentration (0‐2 mM) of aspirin under adipogenic induction medium for 14 d, and the Oil Red O staining was performed, Scale bars, 100 μm. B, Percentage of oil red lipid area was measured. C‐E, The relative expression levels of ap2, PPARγ and C/EBPα in induction group and induction with aspirin group at 3, 7 and 14 d were evaluated by qRT‐PCR. GAPDH was used as an internal control. F, Protein expression of ap2, PPARγ and C/EBPα in induction group and induction with aspirin group at 3, 7 and 14 d was evaluated by Western blot. GAPDH was used as an internal. * P < .05, ** P < .01, *** P < .001, **** P < .000, #: vs. 0.5mM group, N = 3

Article Snippet: We used the following primary antibodies: anti‐ap2 (Proteintech, 1:2000), anti‐C/EBPα (Cell Signaling Technology, 1:2000), anti‐PPARγ (Proteintech, 1:2000), anti‐PTEN (Bioss, 1:2000), anti‐PI3K (Proteintech, 1:2000), anti‐Phospho (P)‐PI3K (Proteintech, 1:2000), anti‐AKT (Bioss, 1:2000) and anti‐P‐AKT (Bioss, 1:2000).

Techniques: Concentration Assay, Staining, Expressing, Quantitative RT-PCR, Control, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Aspirin inhibits adipogenesis of tendon stem cells and lipids accumulation in rat injury tendon through regulating PTEN/PI3K/AKT signalling

doi: 10.1111/jcmm.14622

Figure Lengend Snippet: Effect of aspirin on adipogenic healing. A, Intact tendon, injury tendon and ASA treatment tendon observed through H&E staining. B, Gross appearance of three group tendons. C, Histological score of injury tendon and ASA treatment tendon, N = 9. D‐F, Immunostaining showed the level of ap2 and PPARγ between injury tendon and ASA treatment tendon. ** P < .01, **** P < .000, N = 3

Article Snippet: We used the following primary antibodies: anti‐ap2 (Proteintech, 1:2000), anti‐C/EBPα (Cell Signaling Technology, 1:2000), anti‐PPARγ (Proteintech, 1:2000), anti‐PTEN (Bioss, 1:2000), anti‐PI3K (Proteintech, 1:2000), anti‐Phospho (P)‐PI3K (Proteintech, 1:2000), anti‐AKT (Bioss, 1:2000) and anti‐P‐AKT (Bioss, 1:2000).

Techniques: Staining, Immunostaining