Journal: bioRxiv

Article Title: HDL-AuNPs-BMS nanoparticle conjugates as molecularly targeted therapy for leukemia

doi: 10.1101/250985

Figure Lengend Snippet: HDL-AuNPs-BMS administration in vivo attenuates leukemogenesis. (a) C1498 cells(0.1 × 10 6 ) were intravenously injected into 4–6 weeks old C57BL/6 mice. When the mice showed leukemic disease, free BMS or HDL-AuNPs-BMS was initiated at the dose of 5 mg/kg. A twice-weekly treatment with 5 mg/kg/mouse for 2 weeks, which was continued at the dose of 10 mg/kg/mouse for 1 week while monitoring for any clinical signs of toxicity or leukemia-related distress. (b) Representative visual analysis (left) and calculated weight (right) of spleens from leukemia mice receiving free BMS, HDL-AuNPs-BMS or vehicle. Scale bars, 5.0 mm. (c) External views of lung and liver of untreated and drug-treated leukemia mice. (d) Graph is the quantification of tumor nodules growing in lung or liver. (e,f) H&E staining of tissue sections from lung or liver of leukemia mice receiving free BMS, HDL-AuNPs-BMS or vehicle. Scale bars, 100 μm. (g) Wright-Giemsa-stained cytospins of BM cells from leukemia mice receiving free BMS, HDL-AuNPs-BMS or vehicle (original magnification × 400). Graph is the quantification of post-mitotic cells. (h,j) qPCR analysis of DNMT1 and p15 INK4B expression from BM cells of leukemia mice receiving free BMS, HDL-AuNPs-BMS or vehicle. (i) Dotblot of genomic DNA from BM cells of leukemia mice receiving free BMS, HDL-AuNPs-BMS or vehicle. In all experiments, n = 3 mice/group, data are mean ± SD; * P < 0.05, ** P < 0.01; BMS represents BMS309403; N-BMS represents HDL-AuNPs-BMS. Note: Emp, F-BMS, N-BMS represents empty vehicle, free BMS, HDL-AuNPs-BMS, respectively.

Article Snippet: FABP4 inhibitor (BMS309403) was purchased from Calbiochem and dissolved in ethanol for pre-clinical test and in DMSO for cell culture experiments.

Techniques: In Vivo, Injection, Staining, Expressing

Journal: Oncotarget

Article Title: CRABP-II- and FABP5-independent responsiveness of human glioblastoma cells to all-trans retinoic acid

doi:

Figure Lengend Snippet: the expression patterns of CRABP-II and FABP5 in different grades of human astrocytomas

Article Snippet: FABP5 competitive inhibitor BMS309403 (Santa Cruz) [ , ] was dissolved in DMSO and diluted to the final concentration of 25μM with culture medium.

Techniques: Expressing

Journal: Oncotarget

Article Title: CRABP-II- and FABP5-independent responsiveness of human glioblastoma cells to all-trans retinoic acid

doi:

Figure Lengend Snippet: The staining patterns were scored as “−” if no immunolabeling was observed in the cells and “+” if distinct staining was generally observed. (A) Immunohistochemical profiling of CRABP-II and FABP5 expression patterns in non-tumor brain tissues and glioblastomas. (B) The percentage of CRABP-II and FABP5 expression in the four astrocytoma grades.

Article Snippet: FABP5 competitive inhibitor BMS309403 (Santa Cruz) [ , ] was dissolved in DMSO and diluted to the final concentration of 25μM with culture medium.

Techniques: Staining, Immunolabeling, Immunohistochemical staining, Expressing

Journal: Oncotarget

Article Title: CRABP-II- and FABP5-independent responsiveness of human glioblastoma cells to all-trans retinoic acid

doi:

Figure Lengend Snippet: (A) Immunocytochemical staining of CRABP-II and FABP5 expression in the three cell lines that were either untreated (N) or treated with RA (RA). (B) RT-PCR and western blot analyses of CRABP-II and FABP5 expression in LN18, LN428 and U251 cells with and without RA treatment. CRABP-II/FABP5 ratios were calculated according to the results of RT-PCR and western blotting.

Article Snippet: FABP5 competitive inhibitor BMS309403 (Santa Cruz) [ , ] was dissolved in DMSO and diluted to the final concentration of 25μM with culture medium.

Techniques: Staining, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Oncotarget

Article Title: CRABP-II- and FABP5-independent responsiveness of human glioblastoma cells to all-trans retinoic acid

doi:

Figure Lengend Snippet: (A) RT-PCR examination of CRABP-II, FABP5, RARβ, OLFM4, PPARγ and CYP26A1 transcription under the following conditions: normal culture, 10 μM of RA for 48 h, 1 μM decitabine for 72 h and 10 μM of RA for 48 h after pre-treatment with 1 μM decitabine for 72 h. CRABP-II/FABP5 ratios were calculated according to RT-PCR data. (B) CRABP-II and FABP5 western blotting performed on LN18, LN428 and U251 cells under the following conditions: normal culture, 10 μM of RA for 48 h and 1 μM decitabine pretreatment for 72 h followed by 10 μM of RA for 48 h. CRABP-II/FABP5 ratios were calculated according to the data obtained.

Article Snippet: FABP5 competitive inhibitor BMS309403 (Santa Cruz) [ , ] was dissolved in DMSO and diluted to the final concentration of 25μM with culture medium.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Oncotarget

Article Title: CRABP-II- and FABP5-independent responsiveness of human glioblastoma cells to all-trans retinoic acid

doi:

Figure Lengend Snippet: (A) Hematoxylin and eosin morphological staining performed on LN18, LN428 and U251 cells under the following conditions: normal culture; 25 μM BMS309403 pretreatment for 6 h followed by 10 μM RA treatment for 48 h (BMS+RA) and 10 μM RA treatment for 48 h (RA). (B) Evaluation of the responses by cell counting of LN18, LN428 and U251 treated with 25 μM of BMS309403 for 6 h before 10 μM of RA and 10 μM RA for 72 h. *, compared with N group, p >0.05; #, compared with BMS+RA group, p >0.05.

Article Snippet: FABP5 competitive inhibitor BMS309403 (Santa Cruz) [ , ] was dissolved in DMSO and diluted to the final concentration of 25μM with culture medium.

Techniques: Staining, Cell Counting

Journal: Oncotarget

Article Title: CRABP-II- and FABP5-independent responsiveness of human glioblastoma cells to all-trans retinoic acid

doi:

Figure Lengend Snippet: Primer sequences, amplicon size and annealing temperature for RT-PCR

Article Snippet: FABP5 competitive inhibitor BMS309403 (Santa Cruz) [ , ] was dissolved in DMSO and diluted to the final concentration of 25μM with culture medium.

Techniques: Amplification

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: To examine the effects of rhIL-17A and palmitic acid (PA, Sigma-Aldrich Cat# P9767) on cell proliferation, OVCAR3 and A2780 cells were seeded in 96-well plates at 3 × 10 3 cells per well and were then treated with (1) 0, 0.1, 1, 10, 100 ng/ml rhIL-17A for 12, 24, 48 or 72 h; (2) 0, 12.5, 25, 50, 100 μM PA for 0, 24 or 48 h; (3) 10 ng/ml rhIL-17A and PA (0, 12.5, 25, 50, or 100 μM) for 48 h. To determine the signals involved in the effects of rhIL-17A and PA on cell proliferation, OVCAR3 and A2780 cells were pretreated with the FABP4 inhibitor BMS309403 (25 μM; Merck Cat# 341310) or the STAT3 inhibitor STATTIC (A2780, 0.3125 μM; OVCAR3, 1.25 μM; Selleck Cat# S7024) followed by 48 h of treatment with 10 ng/ml rhIL-17A and 25 μM PA. After treatments, cells were incubated with MTS (KeyGEN BioTECH Cat# KGA327) for an additional 2 h at 37 °C in the dark.

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: To examine the effects of rhIL-17A and palmitic acid (PA, Sigma-Aldrich Cat# P9767) on cell proliferation, OVCAR3 and A2780 cells were seeded in 96-well plates at 3 × 10 3 cells per well and were then treated with (1) 0, 0.1, 1, 10, 100 ng/ml rhIL-17A for 12, 24, 48 or 72 h; (2) 0, 12.5, 25, 50, 100 μM PA for 0, 24 or 48 h; (3) 10 ng/ml rhIL-17A and PA (0, 12.5, 25, 50, or 100 μM) for 48 h. To determine the signals involved in the effects of rhIL-17A and PA on cell proliferation, OVCAR3 and A2780 cells were pretreated with the FABP4 inhibitor BMS309403 (25 μM; Merck Cat# 341310) or the STAT3 inhibitor STATTIC (A2780, 0.3125 μM; OVCAR3, 1.25 μM; Selleck Cat# S7024) followed by 48 h of treatment with 10 ng/ml rhIL-17A and 25 μM PA. After treatments, cells were incubated with MTS (KeyGEN BioTECH Cat# KGA327) for an additional 2 h at 37 °C in the dark.

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: To examine the effects of rhIL-17A and palmitic acid (PA, Sigma-Aldrich Cat# P9767) on cell proliferation, OVCAR3 and A2780 cells were seeded in 96-well plates at 3 × 10 3 cells per well and were then treated with (1) 0, 0.1, 1, 10, 100 ng/ml rhIL-17A for 12, 24, 48 or 72 h; (2) 0, 12.5, 25, 50, 100 μM PA for 0, 24 or 48 h; (3) 10 ng/ml rhIL-17A and PA (0, 12.5, 25, 50, or 100 μM) for 48 h. To determine the signals involved in the effects of rhIL-17A and PA on cell proliferation, OVCAR3 and A2780 cells were pretreated with the FABP4 inhibitor BMS309403 (25 μM; Merck Cat# 341310) or the STAT3 inhibitor STATTIC (A2780, 0.3125 μM; OVCAR3, 1.25 μM; Selleck Cat# S7024) followed by 48 h of treatment with 10 ng/ml rhIL-17A and 25 μM PA. After treatments, cells were incubated with MTS (KeyGEN BioTECH Cat# KGA327) for an additional 2 h at 37 °C in the dark.

Techniques: 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: To examine the effects of rhIL-17A and palmitic acid (PA, Sigma-Aldrich Cat# P9767) on cell proliferation, OVCAR3 and A2780 cells were seeded in 96-well plates at 3 × 10 3 cells per well and were then treated with (1) 0, 0.1, 1, 10, 100 ng/ml rhIL-17A for 12, 24, 48 or 72 h; (2) 0, 12.5, 25, 50, 100 μM PA for 0, 24 or 48 h; (3) 10 ng/ml rhIL-17A and PA (0, 12.5, 25, 50, or 100 μM) for 48 h. To determine the signals involved in the effects of rhIL-17A and PA on cell proliferation, OVCAR3 and A2780 cells were pretreated with the FABP4 inhibitor BMS309403 (25 μM; Merck Cat# 341310) or the STAT3 inhibitor STATTIC (A2780, 0.3125 μM; OVCAR3, 1.25 μM; Selleck Cat# S7024) followed by 48 h of treatment with 10 ng/ml rhIL-17A and 25 μM PA. After treatments, cells were incubated with MTS (KeyGEN BioTECH Cat# KGA327) for an additional 2 h at 37 °C in the dark.

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: To examine the effects of rhIL-17A and palmitic acid (PA, Sigma-Aldrich Cat# P9767) on cell proliferation, OVCAR3 and A2780 cells were seeded in 96-well plates at 3 × 10 3 cells per well and were then treated with (1) 0, 0.1, 1, 10, 100 ng/ml rhIL-17A for 12, 24, 48 or 72 h; (2) 0, 12.5, 25, 50, 100 μM PA for 0, 24 or 48 h; (3) 10 ng/ml rhIL-17A and PA (0, 12.5, 25, 50, or 100 μM) for 48 h. To determine the signals involved in the effects of rhIL-17A and PA on cell proliferation, OVCAR3 and A2780 cells were pretreated with the FABP4 inhibitor BMS309403 (25 μM; Merck Cat# 341310) or the STAT3 inhibitor STATTIC (A2780, 0.3125 μM; OVCAR3, 1.25 μM; Selleck Cat# S7024) followed by 48 h of treatment with 10 ng/ml rhIL-17A and 25 μM PA. After treatments, cells were incubated with MTS (KeyGEN BioTECH Cat# KGA327) for an additional 2 h at 37 °C in the dark.

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: To examine the effects of rhIL-17A and palmitic acid (PA, Sigma-Aldrich Cat# P9767) on cell proliferation, OVCAR3 and A2780 cells were seeded in 96-well plates at 3 × 10 3 cells per well and were then treated with (1) 0, 0.1, 1, 10, 100 ng/ml rhIL-17A for 12, 24, 48 or 72 h; (2) 0, 12.5, 25, 50, 100 μM PA for 0, 24 or 48 h; (3) 10 ng/ml rhIL-17A and PA (0, 12.5, 25, 50, or 100 μM) for 48 h. To determine the signals involved in the effects of rhIL-17A and PA on cell proliferation, OVCAR3 and A2780 cells were pretreated with the FABP4 inhibitor BMS309403 (25 μM; Merck Cat# 341310) or the STAT3 inhibitor STATTIC (A2780, 0.3125 μM; OVCAR3, 1.25 μM; Selleck Cat# S7024) followed by 48 h of treatment with 10 ng/ml rhIL-17A and 25 μM PA. After treatments, cells were incubated with MTS (KeyGEN BioTECH Cat# KGA327) for an additional 2 h at 37 °C in the dark.

Techniques: Expressing

Journal: The Journal of Reproduction and Development

Article Title: FABP4 mediates endoplasmic reticulum stress and autophagy to regulate endometrial epithelial cell function during early sheep gestation

doi: 10.1262/jrd.2023-015

Figure Lengend Snippet: Expression levels of FABP4 in sheep endometrium. (a) FABP4 mRNA expression in sheep endometrium on day 4 and day 15. (b, c) FABP4 protein expression in sheep endometrium on day 4 and 15. The optical density was normalized to the density of β-actin in the same lane. (d, e) Rabbit IgG group was used as the negative control for analyzing the localization and expression of FABP4 in sheep endometrium. Data are presented as mean ± standard error with significant differences at P < 0.05 and extremely significant differences at P < 0.01. * indicates P < 0.05, ** indicates P < 0.01, and no sign indicates that the difference is not significant. The technique was repeated thrice.

Article Snippet: When SEEC growth reached 80–90% confluence, the medium was replaced with fresh DMEM/F12 and treated with 20 μM of the FABP4 small molecule inhibitor BMS309403 (Glpbio, Montclair, NJ, USA, Cas: 300657-03-8) [ ].

Techniques: Expressing, Negative Control

Journal: The Journal of Reproduction and Development

Article Title: FABP4 mediates endoplasmic reticulum stress and autophagy to regulate endometrial epithelial cell function during early sheep gestation

doi: 10.1262/jrd.2023-015

Figure Lengend Snippet: Expression of FABP4 in SEECs. (a) Isolation and purification of SEECs (250 ×). When the cells reached the sixth passage, they became larger and rounder and began to senesce. (b) Immunofluorescence identification of SEECs (CK18) (100 ×). (c) Localization of FABP4 in SEECs (Yellow arrows are FABP4 in the nuclei) (25 ×).

Article Snippet: When SEEC growth reached 80–90% confluence, the medium was replaced with fresh DMEM/F12 and treated with 20 μM of the FABP4 small molecule inhibitor BMS309403 (Glpbio, Montclair, NJ, USA, Cas: 300657-03-8) [ ].

Techniques: Expressing, Isolation, Purification, Immunofluorescence

Journal: The Journal of Reproduction and Development

Article Title: FABP4 mediates endoplasmic reticulum stress and autophagy to regulate endometrial epithelial cell function during early sheep gestation

doi: 10.1262/jrd.2023-015

Figure Lengend Snippet: Hormone treatment of SEECs and detection of changes in FABP4 expression. (a) Protein expression levels of PGR and ER after combined hormone treatment. (b) The mRNA expression levels of ISG15, HOXA10, CXCL10, and RSAD2 after hormone treatment. (c) Levels of the prostaglandin secreted from the SEECs after hormone treatment.(d) Expression levels of FABP4 after hormone treatment. All data are presented as mean ± standard error. Differences were considered significant at P < 0.05 and extremely significant at P < 0.01.

Article Snippet: When SEEC growth reached 80–90% confluence, the medium was replaced with fresh DMEM/F12 and treated with 20 μM of the FABP4 small molecule inhibitor BMS309403 (Glpbio, Montclair, NJ, USA, Cas: 300657-03-8) [ ].

Techniques: Expressing

Journal: The Journal of Reproduction and Development

Article Title: FABP4 mediates endoplasmic reticulum stress and autophagy to regulate endometrial epithelial cell function during early sheep gestation

doi: 10.1262/jrd.2023-015

Figure Lengend Snippet: FABP4 inhibition impedes SEEC function. (a, b) Mobility of SEECs at 0, 24, 48, 72 and 96 h was measured using a scratch test. Migratory capacity was calculated as a percentage of healing area relative to time 0. (c) CCK-8 viable cell counts quantified the proliferative capacity of SEECs treated with hormone and FABP4 inhibitor BMS309403. (d–g) Expression levels of EMT after hormone and inhibitor treatment (E-cadherin, N-cadherin, Vim, and β-catenin). (h, i) Changes in endoplasmic reticulum stress-related protein CHOP and GRP78 were measured. (j, k) Changes in autophagy-related proteins p-mTOR, LC3B II/I, and P62. Data are expressed as mean ± standard error. Differences were considered significant at P < 0.05 and extremely significant at P < 0.01.

Article Snippet: When SEEC growth reached 80–90% confluence, the medium was replaced with fresh DMEM/F12 and treated with 20 μM of the FABP4 small molecule inhibitor BMS309403 (Glpbio, Montclair, NJ, USA, Cas: 300657-03-8) [ ].

Techniques: Inhibition, CCK-8 Assay, Expressing

Journal: The Journal of Reproduction and Development

Article Title: FABP4 mediates endoplasmic reticulum stress and autophagy to regulate endometrial epithelial cell function during early sheep gestation

doi: 10.1262/jrd.2023-015

Figure Lengend Snippet: TG and 3-MA treatment partially restores SEEC function after BMS30940 suppression of FABP4. (a, b) Expression levels of key proteins in the endoplasmic reticulum stress signaling pathway after combined treatment with hormones, BMS309403, and TG. (c, d) Expression levels of key proteins in autophagy and apoptosis signaling pathways after combined treatment with hormones, BMS309403, and 3-MA. (e) Secreted prostaglandin levels from SEECs after combined treatment with hormones, BMS309403, and TG or 3-MA. Data are expressed as mean ± standard error. Differences were considered significant at P < 0.05 and extremely significant at P < 0.01.

Article Snippet: When SEEC growth reached 80–90% confluence, the medium was replaced with fresh DMEM/F12 and treated with 20 μM of the FABP4 small molecule inhibitor BMS309403 (Glpbio, Montclair, NJ, USA, Cas: 300657-03-8) [ ].

Techniques: Expressing, Protein-Protein interactions

Journal: Translational Lung Cancer Research

Article Title: Impaired natural killer cell maturation in lung adenocarcinoma driven by FABP4 and SPON2 downregulation through disrupted lipid metabolism

doi: 10.21037/tlcr-24-1027

Figure Lengend Snippet: Pseudotime ordering and pathway analysis of NK1 cluster. (A) Pseudotime ordering of NK1 reveals a trajectory. (B) Significant shift toward the NK1 phenotype in T group and N group. (C) Differentially expressed genes (rows) along pseudotime (columns) are clustered hierarchically into two modules. Color key from blue to red indicates relative expression from low to high. (D) Expression dynamics of 103 genes in module 2 in pseudotime, shown as pink lines. The red line indicates the average gene expression patterns in module 2. (E) GO analyses of module 2. (F) Superimposition of expression of specific genes ( FABP4 and SPON2 ) on the reprogramming trajectories. FABP4 , fatty acid-binding protein 4; GO, Gene Ontology; N, non-malignant lung; NK, natural killer; SPON2 , spondin2; T, tumor.

Article Snippet: NK92 cells were treated with an FABP4 inhibitor BMS-309403 (20 μmol/L; APExBIO, Houston, TX, USA) for 48 h at 37 °C.

Techniques: Expressing, Gene Expression, Binding Assay

Journal: Translational Lung Cancer Research

Article Title: Impaired natural killer cell maturation in lung adenocarcinoma driven by FABP4 and SPON2 downregulation through disrupted lipid metabolism

doi: 10.21037/tlcr-24-1027

Figure Lengend Snippet: FABP4 and SPON2 expression in lung adenocarcinoma. (A,B) FABP4 and SPON2 expression in NK cells in a pair of representative samples determined using multiplex IF (400×). Scale bar represents 50 μm. (C) Graph bars showing the relative fluorescence intensity of FABP4 and SPON2 expression, measured from 10 NK cells randomly selected per visual field (400×) for each patient. (D) FABP4 and SPON2 expression in patients with lung adenocarcinoma in TCGA. (E,F) FABP4 and SPON2 expression in patient samples determined using IHC (40×). Scale bar represents 20 μm. **, P<0.01; ***, P<0.001. FABP4 , fatty acid-binding protein 4; IHC, immunohistochemistry; ns, not significant; SPON2 , spondin2; TCGA, The Cancer Genome Atlas.

Article Snippet: NK92 cells were treated with an FABP4 inhibitor BMS-309403 (20 μmol/L; APExBIO, Houston, TX, USA) for 48 h at 37 °C.

Techniques: Expressing, Multiplex Assay, Fluorescence, Binding Assay, Immunohistochemistry

Journal: Translational Lung Cancer Research

Article Title: Impaired natural killer cell maturation in lung adenocarcinoma driven by FABP4 and SPON2 downregulation through disrupted lipid metabolism

doi: 10.21037/tlcr-24-1027

Figure Lengend Snippet: Function analysis of FABP4-deficient NK92 cells. (A) Violin plot showing FABP4 , SPON2 , Tbx21 , and EOMES expression in tumors and adjacent tissues in TCGA data. (B) NK92 cells transfected with shFABP4 lentivirus (200×). (C) FABP4 , Tbx21 , and EOMES expression in NK92 cells transfected with shFABP4 analyzed using fluorescence quantitative polymerase chain reaction. (D) Flow cytometry analysis showing the percentage of degranulated NK cells (CD56 + CD107a + ) among total NK cells (CD56 + ). (E) Volcano plot displaying differential expression of lipids in NK cells across various groups. (F,G) Bar and bubble charts illustrating the enriched signaling pathways for these differentially expressed lipids. **, P<0.01; ***, P<0.001. EOMES , eomesodermin; FABP4 , fatty acid-binding protein 4; ns, not significant; SPON2 , spondin2; Tbx21 , T-box 21; TCGA, The Cancer Genome Atlas.

Article Snippet: NK92 cells were treated with an FABP4 inhibitor BMS-309403 (20 μmol/L; APExBIO, Houston, TX, USA) for 48 h at 37 °C.

Techniques: Expressing, Transfection, Fluorescence, Real-time Polymerase Chain Reaction, Flow Cytometry, Quantitative Proteomics, Protein-Protein interactions, Binding Assay