Journal: Frontiers in Immunology

Article Title: Generation and characterization of human induced pluripotent stem cells from neuropathologically confirmed multiple system atrophy patient-derived fibroblasts

doi: 10.3389/fimmu.2026.1641981

Figure Lengend Snippet: Assessment of the cell line identity and the genomic integrity of the six MSA patient-derived iPSCs. (A) Short tandem repeat profiling provided the number of repeating units of 16 polymorphic microsatellite loci, to compare the donor fibroblast cells (reference) and the generated iPSCs and verify their origin. (B) G-banding revealed that the six iPSC lines showed a normal diploid karyotype (n = 46) and matched the sex of the donor patient. (C) Digital PCR allowed us to screen whether specific genomic regions accumulated duplications that may give a strong selective survival or growth advantage to iPSCs. Example of the scatter digital PCR plots displaying simultaneously (1) FAM signals, shown in purple and green to detect the target gene ( ID1 , NCAPD2 , PITX1 , RPS6KB1 , SOAT1 , or STS ), which are located in the 6 most recurrent abnormal regions; and (2) VIM signals, shown in orange and green to detect the reference gene, RPP30 . The ratio between the number of copies of the target gene and of the reference gene was used to determine the normal 2-copy number or copy number variations. The plots were obtained through QuantStudio 3D Digital PCR system. In all cases, we obtained 2-copy number (D) Quantitative Real Time RT-PCR was used for confirming the absence of the Sendai virus in the iPSC lines after 8 passages, through four primer sets (SeV, KOS, Klf4 and c-Myc). Box and Whisker plots depict in log10 scale the gene expression relative to GAPDH gene and normalized to the 7 days post-transduction fibroblasts, calculated by the 2(-ΔΔC(T)) method. In all iPSC clones, the expression levels of the SeV RNA were very low, reaching the limit of detection. Data are shown as the mean ± s.d. n = 3.

Article Snippet: SOAT1 , Hs06606603_cn , FAM , 1q25 , hg38|179355957-179356068 , 111 bp , HaeIII, HindIII, CviQI.

Techniques: Derivative Assay, Generated, Digital PCR, Quantitative RT-PCR, Virus, Whisker Assay, Gene Expression, Transduction, Clone Assay, Expressing

Journal: bioRxiv

Article Title: Novel Role of AcylCoA:cholesterol acyltransferase 1 (ACAT1/SOAT1) in Diabetic Retinopathy

doi: 10.1101/2025.10.09.681484

Figure Lengend Snippet: A. Immunolabeling of retina sections from human donors with DR showed increased ACAT1/SOAT1 immunoreactivity in the photoreceptor outer segments (OS) and retinal pigment epithelial (RPE) cells, as compared with non-diabetic control donors. Donor sample information is provided in supplementary table 1. Scale bar = 50 µm. n=3. B. Vitrectomy tissues collected from patients with PDR showed an increased level of CE, as measured by ELISA. Donor sample information is provided in supplementary table 2. Data are presented as mean ± SEM; n=4-5; **p<0.01.

Article Snippet: Mice were treated with intraperitoneal injections of a specific ACAT1/SOAT1 inhibitor (K604, MedChem Express, 10 mg/Kg in PBS) or vehicle control (PBS) every 2 days.

Techniques: Immunolabeling, Control, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Novel Role of AcylCoA:cholesterol acyltransferase 1 (ACAT1/SOAT1) in Diabetic Retinopathy

doi: 10.1101/2025.10.09.681484

Figure Lengend Snippet: Ten-week old male diabetic Ins2 Akita mice were treated with PBS as vehicle or ACAT1/SOAT1 inhibitor, K604 (10mg/Kg, i.p.), every 2 days for 2 weeks. After 2 weeks of treatment, eyeballs and retinas were collected. A-B. Oil red O staining of frozen sections shows mild reactivity in the ganglion cell layer (GCL) and photoreceptor outer segments (OS) of the Ins2 Akita diabetic retina. Treatment with the ACAT1/SOAT1 inhibitor, K604, reduced this activity. Mean ± SEM; n=4-5; *p<0.05; ns: not significant. C-D. Detection of CE by filipin labeling of frozen retina sections shows prominent increases in CE formation in the ganglion cell (GCL), outer plexiform (OPL) and photoreceptor outer segment (OS) layers of the Ins2 Akita diabetic retina. Treatment with K604 prevented the CE increase. Scale bar = 40 µm; mean ± SEM; n=4; ****p<0.0001. E-F. Total cholesterol and CE levels were measured in retinas and plasma from non-diabetic control mice, Ins2 Akita diabetic mice, and K604-treated Ins2 Akita diabetic mice. This analysis showed upregulation of total cholesterol in plasma of the Ins2 Akita mice but not in retina. However, CE was increased in both retina and plasma of the Ins2 Akita mice. K604 treatment reduced retinal CE formation as well as preventing the increases in both cholesterol and CE in plasma. Data are presented as mean ± SEM; n=4-6; **p<0.01; ***p<0.001; ****p<0.0001; ns: not significant.

Article Snippet: Mice were treated with intraperitoneal injections of a specific ACAT1/SOAT1 inhibitor (K604, MedChem Express, 10 mg/Kg in PBS) or vehicle control (PBS) every 2 days.

Techniques: Staining, Activity Assay, Labeling, Clinical Proteomics, Control

Journal: bioRxiv

Article Title: Novel Role of AcylCoA:cholesterol acyltransferase 1 (ACAT1/SOAT1) in Diabetic Retinopathy

doi: 10.1101/2025.10.09.681484

Figure Lengend Snippet: A. Western blot analysis of retinas from 12-week diabetic Ins2 Akita mice showed marked increases in B. LDLR, C. ACAT1/SOAT1, D. TREM1, E. MCSF, F. VEGF, and G. TNF-α as compared with the WT non-diabetic controls. These increases were normalized by the treatment with K604 (10mg/Kg, ip) every two days from 10 to 12 weeks. Mean ± SEM, n=3-6; *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. H-I . Increased immunoreactivity for ACAT1/SOAT1 was detected in retinal frozen sections from Ins2 Akita diabetic mice, especially in the RGC, OPL, and OS layers. The ACAT1 immunoreactivity was significantly reduced in retinas of Ins2 Akita diabetic mice treated with K604. Data are presented as mean ± SEM; n=3-6; Scale bar = 25 μm.

Article Snippet: Mice were treated with intraperitoneal injections of a specific ACAT1/SOAT1 inhibitor (K604, MedChem Express, 10 mg/Kg in PBS) or vehicle control (PBS) every 2 days.

Techniques: Western Blot

Journal: bioRxiv

Article Title: Novel Role of AcylCoA:cholesterol acyltransferase 1 (ACAT1/SOAT1) in Diabetic Retinopathy

doi: 10.1101/2025.10.09.681484

Figure Lengend Snippet: A-B. DHE imaging showed upregulation of superoxide formation in frozen sections from Ins2 Akita diabetic mice. This effect was blocked by the K604 treatment. n=4. C-D. Immunofluorescent labeling of frozen retina sections with 4-hydroxynonenal (4-HNE) and fluorescence intensity quantification showed increased 4-HNE levels in the ganglion cell layer (GCL), outer plexiform layer (OPL) and photoreceptor outer segments (OS) of the Ins2 Akita mice. K604 treatment blocked this effect. n=4. E-F. Immunolabeling of retina cross-sections showed increased GFAP immunoreactivity in Müller cells of the Ins2 Akita mice, which was blocked in the K604 treated mice. n=4. Scale bar=30 μm. G-K. Quantitative RT-PCR showed upregulation of Acat1/Soat1 , along with proinflammatory mediators – Mcsf1, Il1b, Tnfa , and Nos2 in Ins2 Akita diabetic mouse retinas, compared to non-diabetic controls. K604 treatment significantly inhibited these increases. n=4-6. All the data are presented as mean ± SEM. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

Article Snippet: Mice were treated with intraperitoneal injections of a specific ACAT1/SOAT1 inhibitor (K604, MedChem Express, 10 mg/Kg in PBS) or vehicle control (PBS) every 2 days.

Techniques: Imaging, Labeling, Fluorescence, Immunolabeling, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Novel Role of AcylCoA:cholesterol acyltransferase 1 (ACAT1/SOAT1) in Diabetic Retinopathy

doi: 10.1101/2025.10.09.681484

Figure Lengend Snippet: Ten-week-old male diabetic Ins2 Akita mice were treated with ACAT1/SOAT1 inhibitor, K604 (10 mg/Kg, ip), every 2 days for 2 weeks. After 2 weeks, retinas were collected for analysis A-B. Confocal imaging of retinal flat mounts labeled with the neuronal marker, RBPMS and quantification showed a significant decrease in RPBMS positive retinal ganglion cells in vehicle-treated Ins2 Akita diabetic retinas compared to the non-diabetic control retinas. This retinal ganglion cell loss was markedly reduced by the K604 treatment. n=5-6; scale bar=40 μm. Data are presented as mean ± SEM. C-D . Retinal function was measured by electroretinography (ERG). Both scotopic a-wave and scotopic b-wave were improved in K604-treated Ins2 Akita diabetic mice as compared with the vehicle treated diabetic mice. n=12–26. Data are presented as mean ± SEM. **p<0.01 non-diabetic versus vehicle-treated diabetic mice; p<0.01 vehicle-treated diabetic versus K604-treated diabetic mice. E . Measurement of visual acuity by optoMotry responses (OMR) showed a marked improvement in the K604-treated diabetic mice as compared with the vehicle-treated diabetic mice. n=10-13. Data are presented as mean ± SEM. *p<0.05; ***p<0.0001.

Article Snippet: Mice were treated with intraperitoneal injections of a specific ACAT1/SOAT1 inhibitor (K604, MedChem Express, 10 mg/Kg in PBS) or vehicle control (PBS) every 2 days.

Techniques: Imaging, Labeling, Marker, Control

Journal: bioRxiv

Article Title: Novel Role of AcylCoA:cholesterol acyltransferase 1 (ACAT1/SOAT1) in Diabetic Retinopathy

doi: 10.1101/2025.10.09.681484

Figure Lengend Snippet: Ten week old male diabetic Ins2 Akita mice were treated with ACAT1/SOAT1 inhibitor, K604 (10 mg/Kg, ip), every 2 days for 2 weeks. After the 2 weeks treatment, retinas were collected for analysis. A-B. Quantitative analysis shows diabetes-induced increases in leukocyte adhesion in Ins2 Akita retinal vessels, which were significantly prevented by K604 treatment as compared with vehicles. Data are presented as mean ± SEM. n=5-6; *p<0.05; **p<0.01; Scale bar=50 μm. C-D . Vascular leakage was visualized and quantified by measuring the fluorescence intensity of FITC-conjugated BSA. K604 treatment significantly prevented vascular leakage in Ins2 Akita mice as compared with vehicle controls. Data are presented as mean ± SEM. n=5-6; scale bar=300 μm; ****p<0.0001. E . K604 treatment significantly prevented diabetes-induced increases in acellular capillary formation as compared with the vehicle controls. Data are presented as mean ± SEM. n=6, scale bar=40 μm; ****p<0.001.

Article Snippet: Mice were treated with intraperitoneal injections of a specific ACAT1/SOAT1 inhibitor (K604, MedChem Express, 10 mg/Kg in PBS) or vehicle control (PBS) every 2 days.

Techniques: Fluorescence

Journal: bioRxiv

Article Title: Novel Role of AcylCoA:cholesterol acyltransferase 1 (ACAT1/SOAT1) in Diabetic Retinopathy

doi: 10.1101/2025.10.09.681484

Figure Lengend Snippet: Eight-months-old male Ins2 Akita diabetic mice were treated with ACAT1/SOAT1 inhibitor, K604 (10 mg/Kg, ip), every 2 days for 2 months. After the 2 months treatment, retinas were collected for analysis. Oil red O staining ( A, B ), filipin staining ( C, D ) and 4HNE staining ( E, F ) of frozen sections shows a strong reaction in areas of the ganglion cell layer (GCL) and photoreceptor outer segments (OS) of the DR retina, indicating lipid accumulation, CE formation, and oxidative stress, respectively. Treatment with the ACAT1/SOAT1 inhibitor, K604, significantly blocked these effects. Scale bar=25 μm. Data are presented as mean ± SEM; n=3-4; **p<0.01; ***p<0.001; ****p<0.0001. G-J. Western blot analysis of retinal samples from the Ins2 Akita diabetic mice showed marked increases in LDLR, ACAT1/SOAT1, and TREM1, as compared with the WT non-diabetic controls. These increases were normalized by treatment with K604. Mean ± SEM; n=3; *p<0.05; ***p<0.001; ****p<0.0001.

Article Snippet: Mice were treated with intraperitoneal injections of a specific ACAT1/SOAT1 inhibitor (K604, MedChem Express, 10 mg/Kg in PBS) or vehicle control (PBS) every 2 days.

Techniques: Staining, Western Blot

Journal: bioRxiv

Article Title: Novel Role of AcylCoA:cholesterol acyltransferase 1 (ACAT1/SOAT1) in Diabetic Retinopathy

doi: 10.1101/2025.10.09.681484

Figure Lengend Snippet: Eight-months old male Ins2 Akita diabetic mice were treated with ACAT1/SOAT1 inhibitor, K604 (10 mg/Kg, ip), every 2 days for 2 months. After 2 months, retinas were collected for analysis. A-B . Albumin staining of retina sections showed a strong reaction in areas of Isolectin B4-positive blood vessels in the diabetic retina, indicating vascular leakage. Treatment with ACAT1/SOAT1 inhibitor, K604, significantly blocked this effect. Scale bar=25 μm; data are presented as mean ± SEM; n=3-4; *p<0.05; ****p<0.0001. C . Measurement of visual acuity by optoMotry responses (OMR) showed a significant impairment in the vehicle-treated diabetic Ins2 Akita mice that was significantly blunted in the K604-treated diabetic mice. n=6-10. Data are presented as mean ± SEM; **p<0.01; ****p<0.0001. D-E . Electroretinography (ERG) showed improvement in both scotopic a-wave and scotopic b-wave in K604-treated diabetic mice as compared with the vehicle treated diabetic mice. n=6-10. Data are presented as mean ± SEM. **p<0.01 non-diabetic versus vehicle-treated diabetic mice; p<0.01 vehicle-treated diabetic versus K604-treated diabetic mice.

Article Snippet: Mice were treated with intraperitoneal injections of a specific ACAT1/SOAT1 inhibitor (K604, MedChem Express, 10 mg/Kg in PBS) or vehicle control (PBS) every 2 days.

Techniques: Staining

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A Experimental design of an in vivo high-throughput screening for anti-tumor immunotherapeutic agents using Drosophila stem cell-derived tumor model. Created in BioRender. Lee, H. (2026) https://BioRender.com/4zb6pi7 . B Representative images showing the immunofluorescent staining of GFP + Arm + Ras V12 -transformed renal nephric stem cells in Drosophila treated with DMSO or STK. C Quantification of the tumor size of renal nephric stem cell tumors in ( B ) ( n = 12 biologically independent samples). D Experimental design of strategies to identify target proteins of STK. The reverse virtual screening was performed by using Pharmmapper and Chemmapper databases for pharmacophore matching and potential target identification, followed by Drug Affinity Responsive Target Stability (DARTS) experiments. Created in BioRender. Lee, H. (2026) https://BioRender.com/lb2oxp7 . E An Venn diagram of potential target proteins identified by virtual screening and DARTS. F Immunoblotting analysis of DARTS detecting the binding of STK to SOAT1. Data are representative of three independent experiments ( n = 3 biological replicates). G Molecular docking analysis of STK binding to human SOAT1. H Isothermal titration calorimetry (ITC) analysis of binding of STK to SOAT1 purified from E. coli . I , J Tumor growth curves and tumor weight of BALB/c mice transplanted with Scram-knockdown or Soat1 -knockdown CT26 cells ( n = 5 biologically independent samples). Data are shown as the mean ± SD. Scale bars are as indicated. Statistical significance was assessed by a two-tailed unpaired Student’s t -test or two-way ANOVA. Source data are provided as a Source Data file.

Article Snippet: L – N SOAT1 protein expression profiles in paired normal and tumor tissues across human lung, colorectal and hepatic carcinomas derived from Human Protein Atlas (HPA) database ( n = 3 biological replicates).

Techniques: In Vivo, High Throughput Screening Assay, Derivative Assay, Staining, Transformation Assay, Drug discovery, Western Blot, Binding Assay, Isothermal Titration Calorimetry, Purification, Knockdown, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A Schematic diagram of treatment design for Lgr5-CreER-GFP/Apc f/f mice. Created in BioRender. Lee, H. (2026) https://BioRender.com/ol3f47g . B Representative images of immunohistochemistry staining of cancer stem cells in the intestine of CSC-like colon cancer mice treated with DMSO or STK ( n = 12 mice in each group). C Quantification of tumor numbers in ( B ) ( n = 12 biologically independent samples). D Schematic diagram of treatment design for MYC-ON mice. Created in BioRender. Lee, H. (2026) https://BioRender.com/0wx33vy . E Representative liver images of MYC-ON mice treated with DMSO (left) or STK (right). F Tumor numbers of MYC-ON mice treated with DMSO or STK. ( n = 8 biologically independent samples). G Survival curves of MYC-ON mice treated with DMSO or STK. ( n = 6 biologically independent samples). H Schematic diagram of treatment design for LSL-Kras/P53 f/f mice. Created in BioRender. Lee, H. (2026) https://BioRender.com/kh1x47x . I Representative lung images of lung adenocarcinoma mice treated with DMSO (left) or STK (right). J Tumor numbers of lung cancer mice treated with DMSO or STK. ( n = 10 mice in each group). K Survial curves of lung cancer mice treated with DMSO or STK. ( n =8 mice in each group). L – N SOAT1 protein expression profiles in paired normal and tumor tissues across human lung, colorectal and hepatic carcinomas derived from Human Protein Atlas (HPA) database ( n = 3 biological replicates). Images shown are representative results from multiple tissue samples. O – S Kaplan–Meier analysis of the correlation between SOAT1 expression levels and overall survival (OS) of patients from the TCGA dataset. T Immunoblotting analysis of SOAT1 levels in the stem CT26 subpopulation and non-stem CT26 subpopulation. U Flow cytometry analysis of SOAT1 expression levels between normal and tumor tissues from human lung carcinoma patients. V Flow cytometry analysis of SOAT1 expression levels on stem cancer cells or non-stem cancer cells from human lung carcinoma patients. W Immunofluorescence (IF) staining of tumor tissues from CT26 subcutaneous tumor-bearing mice ( n = 3 biological replicates). Orange: CD133; Green: SOAT1; Blue: DAPI. Yellow arrowheads indicate cells with co-localization of CD133 and SOAT1. Scale bar: 10 µm. Shown is a representative image from three independent experiments. Data are shown as the mean ± SD or the Kaplan-Meier test. Scale bars are as indicated. Statistical significance was assessed by a two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file.

Article Snippet: L – N SOAT1 protein expression profiles in paired normal and tumor tissues across human lung, colorectal and hepatic carcinomas derived from Human Protein Atlas (HPA) database ( n = 3 biological replicates).

Techniques: Immunohistochemistry, Staining, Expressing, Derivative Assay, Western Blot, Flow Cytometry, Immunofluorescence, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A Volcano plot of differentially expressed genes in tumor-infiltrated Tregs from live cancer mice treated with or without STK. B NicheNet ligand-receptor analysis plot between tumor cells and Tregs from lung cancer mice treated with or without STK. C Representative flow cytometry plots of GPR132, CTLA-4 and LAG3 subpopulation in Tregs in LSL-Kras/P53 f/f /Soat1 f/f mice or LSL-Kras/P53 f/f mice. D – F Quantification of GPR132, CTLA-4 and LAG-3 positive percentage in LSL-Kras/P53 f/f /Soat1 f/f mice or LSL-Kras/P53 f/f mice. ( n = 3 in each group) ( G , H ) Kaplan–Meier analysis of the correlation between GPR132, FOXP3 and CTLA-4 expression levels and overall survival (OS)/disease-free survival (DFS) in patients from the TCGA dataset. I , J Tumor growth curves of BALB/c mice transplanted with CT26 cells with or without anti-CD25 antibody treatment, in combination with DMSO or STK. ( n = 5 biologically independent samples). K – N Tumor growth curves and tumor weight in Foxp3 Cre mice (K and L) or Foxp3 Cre ; Gpr132 f/f mice (M and N). LLC cells transfected with ShScram or Sh Soat1 were transplanted into Foxp3 Cre or Foxp3 Cre ; Gpr132 f/f mice ( n = 5 biologically independent samples). Data are shown as the mean ± SD. Statistical significance was assessed by a two-tailed unpaired Student’s t -test or two-way ANOVA. Source data are provided as a Source Data file.

Article Snippet: L – N SOAT1 protein expression profiles in paired normal and tumor tissues across human lung, colorectal and hepatic carcinomas derived from Human Protein Atlas (HPA) database ( n = 3 biological replicates).

Techniques: Flow Cytometry, Expressing, Transfection, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A , B Heatmap of pathway enrichment analysis of tumor-infiltrated Tregs from liver cancer mice treated with or without STK, based on scRNA-seq data. C Proposed model of GPR132 signaling pathway in Tregs. Created in BioRender. Lee, H. (2026) https://BioRender.com/ou98v3q . D , F Secretion levels of TGF-β and IL-10 in Tregs cultured with conditional medium (CM) from DMSO-treated or STK-treated CT26 cells. ( n = 13 or 9 in each group). E , G Secretion levels of TGF-β and IL-10 in Tregs cultured with conditioned medium from the Scramble -knockdown or Soat1 -knockdown CT26 cells. ( n = 6 in each group). H , I Secretion levels of TGF-β in Tregs cultured with conditioned medium from indicated cells treated with GPR132 inhibitor telmisartan or STAT1 inhibitor fludarabine. ( n = 3 or 4 in each group). J , K Secretion levels of IL-10 in Tregs cultured with conditioned medium from indicated cells treated with GPR132 inhibitor telmisartan or STAT1 inhibitor fludarabine. ( n = 3 or 4 in each group). L – O Expression of GPR132 and LAG-3 on Tregs cultured in CM from STK or DMSO-treated CT26 cells. Tregs were concurrently treated with vehicle, GPR132 inhibitor telmisartan or STAT1 inhibitor fludarabine. Representative flow cytometry plots of GPR132(L) and LAG-3(N). Quantification of GPR132 (M) and LAG-3 (O) in Treg cells. ( n = 4 in each group). Data are shown as the mean ± SD. Statistical significance was assessed by a two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file.

Article Snippet: L – N SOAT1 protein expression profiles in paired normal and tumor tissues across human lung, colorectal and hepatic carcinomas derived from Human Protein Atlas (HPA) database ( n = 3 biological replicates).

Techniques: Cell Culture, Knockdown, Expressing, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A , B Flow cytometry analysis and statistical analysis of NBD-labeled cholesterol uptake. Tregs were cultured with conditioned medium (CM) collected from CT26 cells that had been pulsed with NBD-cholesterol and treated with DMSO or STK. ( n = 5 in each group). C Enrichment of gene sets related to oxysterol metabolism in tumor cells based on proteomic data. D Heatmap of oxysterol-related enzymes mRNA levels of bulk RNA sequencing from Hepa1-6 cells. ( n = 3 in each group). E Heatmap analysis of oxysterol-related enzymes mRNA levels in Hepa1-6 cells treated with DMSO or STK detected by qPCR. ( n = 3 in each group). F Heatmap analysis of mRNA of oxysterol-related enzymes in CT26 cells with Soat1 knockdown or treated with STK detected by qPCR. ( n = 3 in each group). G , H LC-MS analysis of oxysterols in the conditioned medium of Hepa1-6 or CT26 cells treated with DMSO or STK. I Immunoblotting analysis of DARTS-based detection for the binding of 20SOHC to GPR132. Data are representative of three independent experiments. J , K Molecular docking analysis of 20SOHC or 9S-HODE binding to human GPR132. Data are shown as the mean ± SD. Statistical significance was assessed by a two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file.

Article Snippet: L – N SOAT1 protein expression profiles in paired normal and tumor tissues across human lung, colorectal and hepatic carcinomas derived from Human Protein Atlas (HPA) database ( n = 3 biological replicates).

Techniques: Flow Cytometry, Labeling, Cell Culture, RNA Sequencing, Knockdown, Liquid Chromatography with Mass Spectroscopy, Western Blot, Binding Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A , D Representative tumor images of CT26 syngeneic tumor-bearing mice or 4T1 syngeneic tumor-bearing mice treated with DMSO or STK, in combination with anti-PD-1 blocking monoclonal antibodies (mAb). B , C Tumor growth curves and tumor weight of CT26 syngeneic tumor-bearing mice with indicated treatments ( n = 6 biologically independent samples). E , F Tumor growth curves and tumor weight of 4T1 syngeneic cancer mice with indicated treatments ( n = 5 biologically independent samples). G Experimental design of generating patient-derived xenograft (PDX) models of colon cancer in humanized immune system mice. Created in BioRender. Lee, H. (2026) https://BioRender.com/eu4e0mu . H , I Tumor growth curves and tumor weight of PDX mice with indicated treatment ( n = 6 biologically independent samples). J – M Flow cytometry analysis of the proportion of Tregs, the levels of GPR132 and CTLA4 in Tregs, and the proportion of CD8 + T cells in PDX mice ( n = 3 biologically independent samples). N Flow cytometry analysis of GPR132 expression in Tregs from paired lung carcinoma patient tumor samples and normal tissues. O The correlation analysis of SOAT1 expression in tumor cells and GPR132 expression levels in Tregs detected in lung carcinoma patient tumor samples. P , Q Kaplan–Meier analysis of the correlation between Cyp11a1 levels and overall survival (OS) in liver cancer patients from the TCGA dataset. Data are shown as the mean ± SD. Scale bars are as indicated. Statistical significance was assessed by a two-way ANOVA or two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file.

Article Snippet: L – N SOAT1 protein expression profiles in paired normal and tumor tissues across human lung, colorectal and hepatic carcinomas derived from Human Protein Atlas (HPA) database ( n = 3 biological replicates).

Techniques: Blocking Assay, Bioprocessing, Derivative Assay, Flow Cytometry, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A Experimental design of an in vivo high-throughput screening for anti-tumor immunotherapeutic agents using Drosophila stem cell-derived tumor model. Created in BioRender. Lee, H. (2026) https://BioRender.com/4zb6pi7 . B Representative images showing the immunofluorescent staining of GFP + Arm + Ras V12 -transformed renal nephric stem cells in Drosophila treated with DMSO or STK. C Quantification of the tumor size of renal nephric stem cell tumors in ( B ) ( n = 12 biologically independent samples). D Experimental design of strategies to identify target proteins of STK. The reverse virtual screening was performed by using Pharmmapper and Chemmapper databases for pharmacophore matching and potential target identification, followed by Drug Affinity Responsive Target Stability (DARTS) experiments. Created in BioRender. Lee, H. (2026) https://BioRender.com/lb2oxp7 . E An Venn diagram of potential target proteins identified by virtual screening and DARTS. F Immunoblotting analysis of DARTS detecting the binding of STK to SOAT1. Data are representative of three independent experiments ( n = 3 biological replicates). G Molecular docking analysis of STK binding to human SOAT1. H Isothermal titration calorimetry (ITC) analysis of binding of STK to SOAT1 purified from E. coli . I , J Tumor growth curves and tumor weight of BALB/c mice transplanted with Scram-knockdown or Soat1 -knockdown CT26 cells ( n = 5 biologically independent samples). Data are shown as the mean ± SD. Scale bars are as indicated. Statistical significance was assessed by a two-tailed unpaired Student’s t -test or two-way ANOVA. Source data are provided as a Source Data file.

Article Snippet: We further investigated the expression of SOAT1 protein in human lung, colon, and liver cancers using the Human Protein Atlas (HPA) database.

Techniques: In Vivo, High Throughput Screening Assay, Derivative Assay, Staining, Transformation Assay, Drug discovery, Western Blot, Binding Assay, Isothermal Titration Calorimetry, Purification, Knockdown, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A Schematic diagram of treatment design for Lgr5-CreER-GFP/Apc f/f mice. Created in BioRender. Lee, H. (2026) https://BioRender.com/ol3f47g . B Representative images of immunohistochemistry staining of cancer stem cells in the intestine of CSC-like colon cancer mice treated with DMSO or STK ( n = 12 mice in each group). C Quantification of tumor numbers in ( B ) ( n = 12 biologically independent samples). D Schematic diagram of treatment design for MYC-ON mice. Created in BioRender. Lee, H. (2026) https://BioRender.com/0wx33vy . E Representative liver images of MYC-ON mice treated with DMSO (left) or STK (right). F Tumor numbers of MYC-ON mice treated with DMSO or STK. ( n = 8 biologically independent samples). G Survival curves of MYC-ON mice treated with DMSO or STK. ( n = 6 biologically independent samples). H Schematic diagram of treatment design for LSL-Kras/P53 f/f mice. Created in BioRender. Lee, H. (2026) https://BioRender.com/kh1x47x . I Representative lung images of lung adenocarcinoma mice treated with DMSO (left) or STK (right). J Tumor numbers of lung cancer mice treated with DMSO or STK. ( n = 10 mice in each group). K Survial curves of lung cancer mice treated with DMSO or STK. ( n =8 mice in each group). L – N SOAT1 protein expression profiles in paired normal and tumor tissues across human lung, colorectal and hepatic carcinomas derived from Human Protein Atlas (HPA) database ( n = 3 biological replicates). Images shown are representative results from multiple tissue samples. O – S Kaplan–Meier analysis of the correlation between SOAT1 expression levels and overall survival (OS) of patients from the TCGA dataset. T Immunoblotting analysis of SOAT1 levels in the stem CT26 subpopulation and non-stem CT26 subpopulation. U Flow cytometry analysis of SOAT1 expression levels between normal and tumor tissues from human lung carcinoma patients. V Flow cytometry analysis of SOAT1 expression levels on stem cancer cells or non-stem cancer cells from human lung carcinoma patients. W Immunofluorescence (IF) staining of tumor tissues from CT26 subcutaneous tumor-bearing mice ( n = 3 biological replicates). Orange: CD133; Green: SOAT1; Blue: DAPI. Yellow arrowheads indicate cells with co-localization of CD133 and SOAT1. Scale bar: 10 µm. Shown is a representative image from three independent experiments. Data are shown as the mean ± SD or the Kaplan-Meier test. Scale bars are as indicated. Statistical significance was assessed by a two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file.

Article Snippet: We further investigated the expression of SOAT1 protein in human lung, colon, and liver cancers using the Human Protein Atlas (HPA) database.

Techniques: Immunohistochemistry, Staining, Expressing, Derivative Assay, Western Blot, Flow Cytometry, Immunofluorescence, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A Volcano plot of differentially expressed genes in tumor-infiltrated Tregs from live cancer mice treated with or without STK. B NicheNet ligand-receptor analysis plot between tumor cells and Tregs from lung cancer mice treated with or without STK. C Representative flow cytometry plots of GPR132, CTLA-4 and LAG3 subpopulation in Tregs in LSL-Kras/P53 f/f /Soat1 f/f mice or LSL-Kras/P53 f/f mice. D – F Quantification of GPR132, CTLA-4 and LAG-3 positive percentage in LSL-Kras/P53 f/f /Soat1 f/f mice or LSL-Kras/P53 f/f mice. ( n = 3 in each group) ( G , H ) Kaplan–Meier analysis of the correlation between GPR132, FOXP3 and CTLA-4 expression levels and overall survival (OS)/disease-free survival (DFS) in patients from the TCGA dataset. I , J Tumor growth curves of BALB/c mice transplanted with CT26 cells with or without anti-CD25 antibody treatment, in combination with DMSO or STK. ( n = 5 biologically independent samples). K – N Tumor growth curves and tumor weight in Foxp3 Cre mice (K and L) or Foxp3 Cre ; Gpr132 f/f mice (M and N). LLC cells transfected with ShScram or Sh Soat1 were transplanted into Foxp3 Cre or Foxp3 Cre ; Gpr132 f/f mice ( n = 5 biologically independent samples). Data are shown as the mean ± SD. Statistical significance was assessed by a two-tailed unpaired Student’s t -test or two-way ANOVA. Source data are provided as a Source Data file.

Article Snippet: We further investigated the expression of SOAT1 protein in human lung, colon, and liver cancers using the Human Protein Atlas (HPA) database.

Techniques: Flow Cytometry, Expressing, Transfection, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A , B Heatmap of pathway enrichment analysis of tumor-infiltrated Tregs from liver cancer mice treated with or without STK, based on scRNA-seq data. C Proposed model of GPR132 signaling pathway in Tregs. Created in BioRender. Lee, H. (2026) https://BioRender.com/ou98v3q . D , F Secretion levels of TGF-β and IL-10 in Tregs cultured with conditional medium (CM) from DMSO-treated or STK-treated CT26 cells. ( n = 13 or 9 in each group). E , G Secretion levels of TGF-β and IL-10 in Tregs cultured with conditioned medium from the Scramble -knockdown or Soat1 -knockdown CT26 cells. ( n = 6 in each group). H , I Secretion levels of TGF-β in Tregs cultured with conditioned medium from indicated cells treated with GPR132 inhibitor telmisartan or STAT1 inhibitor fludarabine. ( n = 3 or 4 in each group). J , K Secretion levels of IL-10 in Tregs cultured with conditioned medium from indicated cells treated with GPR132 inhibitor telmisartan or STAT1 inhibitor fludarabine. ( n = 3 or 4 in each group). L – O Expression of GPR132 and LAG-3 on Tregs cultured in CM from STK or DMSO-treated CT26 cells. Tregs were concurrently treated with vehicle, GPR132 inhibitor telmisartan or STAT1 inhibitor fludarabine. Representative flow cytometry plots of GPR132(L) and LAG-3(N). Quantification of GPR132 (M) and LAG-3 (O) in Treg cells. ( n = 4 in each group). Data are shown as the mean ± SD. Statistical significance was assessed by a two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file.

Article Snippet: We further investigated the expression of SOAT1 protein in human lung, colon, and liver cancers using the Human Protein Atlas (HPA) database.

Techniques: Cell Culture, Knockdown, Expressing, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A , B Flow cytometry analysis and statistical analysis of NBD-labeled cholesterol uptake. Tregs were cultured with conditioned medium (CM) collected from CT26 cells that had been pulsed with NBD-cholesterol and treated with DMSO or STK. ( n = 5 in each group). C Enrichment of gene sets related to oxysterol metabolism in tumor cells based on proteomic data. D Heatmap of oxysterol-related enzymes mRNA levels of bulk RNA sequencing from Hepa1-6 cells. ( n = 3 in each group). E Heatmap analysis of oxysterol-related enzymes mRNA levels in Hepa1-6 cells treated with DMSO or STK detected by qPCR. ( n = 3 in each group). F Heatmap analysis of mRNA of oxysterol-related enzymes in CT26 cells with Soat1 knockdown or treated with STK detected by qPCR. ( n = 3 in each group). G , H LC-MS analysis of oxysterols in the conditioned medium of Hepa1-6 or CT26 cells treated with DMSO or STK. I Immunoblotting analysis of DARTS-based detection for the binding of 20SOHC to GPR132. Data are representative of three independent experiments. J , K Molecular docking analysis of 20SOHC or 9S-HODE binding to human GPR132. Data are shown as the mean ± SD. Statistical significance was assessed by a two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file.

Article Snippet: We further investigated the expression of SOAT1 protein in human lung, colon, and liver cancers using the Human Protein Atlas (HPA) database.

Techniques: Flow Cytometry, Labeling, Cell Culture, RNA Sequencing, Knockdown, Liquid Chromatography with Mass Spectroscopy, Western Blot, Binding Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibition of the cancer stem cell immune checkpoint SOAT1 suppresses regulatory T cell functions through a trans-cellular 20(S)-Hydroxycholesterol-GPR132 pathway in mice

doi: 10.1038/s41467-026-69305-3

Figure Lengend Snippet: A , D Representative tumor images of CT26 syngeneic tumor-bearing mice or 4T1 syngeneic tumor-bearing mice treated with DMSO or STK, in combination with anti-PD-1 blocking monoclonal antibodies (mAb). B , C Tumor growth curves and tumor weight of CT26 syngeneic tumor-bearing mice with indicated treatments ( n = 6 biologically independent samples). E , F Tumor growth curves and tumor weight of 4T1 syngeneic cancer mice with indicated treatments ( n = 5 biologically independent samples). G Experimental design of generating patient-derived xenograft (PDX) models of colon cancer in humanized immune system mice. Created in BioRender. Lee, H. (2026) https://BioRender.com/eu4e0mu . H , I Tumor growth curves and tumor weight of PDX mice with indicated treatment ( n = 6 biologically independent samples). J – M Flow cytometry analysis of the proportion of Tregs, the levels of GPR132 and CTLA4 in Tregs, and the proportion of CD8 + T cells in PDX mice ( n = 3 biologically independent samples). N Flow cytometry analysis of GPR132 expression in Tregs from paired lung carcinoma patient tumor samples and normal tissues. O The correlation analysis of SOAT1 expression in tumor cells and GPR132 expression levels in Tregs detected in lung carcinoma patient tumor samples. P , Q Kaplan–Meier analysis of the correlation between Cyp11a1 levels and overall survival (OS) in liver cancer patients from the TCGA dataset. Data are shown as the mean ± SD. Scale bars are as indicated. Statistical significance was assessed by a two-way ANOVA or two-tailed unpaired Student’s t -test. Source data are provided as a Source Data file.

Article Snippet: We further investigated the expression of SOAT1 protein in human lung, colon, and liver cancers using the Human Protein Atlas (HPA) database.

Techniques: Blocking Assay, Bioprocessing, Derivative Assay, Flow Cytometry, Expressing, Two Tailed Test

Journal: Frontiers in Nutrition

Article Title: Long-term dietary restriction changes lipid homeostasis and consequently impairs testosterone production in aged Wistar rats

doi: 10.3389/fnut.2025.1665682

Figure Lengend Snippet: LTDR changed expression of the genes involved in lipid metabolism in the liver. To assess the effects of LTDR on gene expression, pieces of the livers from the adult, aged and LTDR groups were collected, total RNA was extracted, and qPCR was performed. Target gene expression (%) was determined by the comparative 2 −ΔΔCT quantification method with Gapdh as the reference gene. Time- and LTDR-induced changes in expression profile of Hmgcr , Srebf1 , Soat1 , Lxra , ApoB , Ldlr , ApoA , Scarb1 , Npc1 , Lrp , Abca1 are presented at panels A–K . The bars shown are mean ± SEM values; for all the analysis n ≥ 5; * indicates significant differences compared to adults; # indicates significant differences between aged animals and the LTDR group; p < 0.05.

Article Snippet: Used Applied Biosystems predesigned TaqMan Gene Expression Assays: Abca1 (Rn00710172_m1), Apob (01499054_m1), Cyp46a1 (Rn01430187_g1), Gapdh (Rn01462662_g1), Hmgc (Rn 00565598_m1), Lrp1 (Rn01503901), Ldlr (Rn00598442_m1), Npc1 (Rn01531821_m1), Soat1 (Rn_00579605_m1), Srebf1 (Rn01495769_m1); sequences of used forward and reverse primers are listed in .

Techniques: Expressing, Gene Expression, Targeted Gene Expression