Journal: Science Advances

Article Title: Shifting IRES versus Cap-initiated translation during homeostatic stem cell differentiation and stress

doi: 10.1126/sciadv.adz7896

Figure Lengend Snippet: ( A ) Representative gating strategy for sorting low, mid, and high IRES/Cap SLAM-LKS. ( B and C ) Colony formation (B) and size (C, mm 2 ) of Translator SLAM-LKS sorted on the basis of IRES/Cap ( n = 6). ( D and E ) Colony formation (D) and size (E, mm 2 ) of Translator GMP sorted on the basis of IRES/Cap ( n = 9). ( F ) Total protein synthesis rates of Translator GMP based on IRES/Cap measured by AF647-OPP MFI ( n = 3). ( G ) Colony formation of EMCV-IRES reporter-transduced CD34+CD90+EPCR+CD45RA− human cord blood sorted on the basis of IRES/Cap ( n = 3). ( H and I ) Colony formation of SLAM-LKS (H) and GMPs (I) based on RUNX1-IRES/Cap sorted from transplant recipients ( n = 3 and 2, respectively). ( J ) Primary competitive transplant schema. Twenty CD45.2 SLAM-LKS sorted on the basis of IRES/Cap versus 200,000 CD45.1 whole BM cells in lethally irradiated CD45.1 recipients. Peripheral blood chimerism analyzed every four weeks through week 16. Created in BioRender. Li, D. (2026) https://BioRender.com/d7k2abt . ( K ) Total chimerism Translator SLAM-LKS based on IRES/Cap in primary recipients evaluated through week 16 ( n = 12). ( L ) Secondary competitive transplant schema. One million total BM cells were collected from primary recipients (16 weeks posttransplantation) and transplanted to lethally irradiated CD45.1 recipients. Created in BioRender. Li, D. (2026) https://BioRender.com/d7k2abt . ( M ) Total chimerism Translator SLAM-LKS based on IRES/Cap in secondary recipients through week 16 ( n = 12). ( N and O ) Representative image of the culture of a single low (N) or high (O) IRES/Cap SLAM-LKS after 5 days of culture. ( P and Q ) Pie charts depicting the percentage of wells with megakaryocytes (MK) after 5 days of culturing a single low (P) or high (Q) IRES/Cap Translator SLAM-LKS ( n = 20). Data show individual replicates and means ± SEM. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001. Significance was assessed using a two-way ANOVA with Dunnett’s one-way ANOVA (B to I) or an ANOVA with Tukey’s multiple comparisons test (K and M).

Article Snippet: WT CD45.2 C57BL/6J (strain #000664), CD45.1 (B6.SJL- Ptprc a Pepc b /BoyJ; strain #002014), KH2/iCas9 [B6;129S4- Gt(ROSA)26Sor tm1(rtTA*M2)Jae Col1a1 tm1(tetO-cas9)Sho /J; strain #029415], Rpl24 Bst (C57BLKS-Rpl24 Bst /J; strain #000516), and NSG mice (NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ; strain #005557) were purchased from the Jackson Laboratory.

Techniques: Irradiation

Journal: Science Advances

Article Title: Shifting IRES versus Cap-initiated translation during homeostatic stem cell differentiation and stress

doi: 10.1126/sciadv.adz7896

Figure Lengend Snippet: ( A ) Representative gating strategy for sorting low, mid, and high IRES/Cap SLAM-LKS. ( B and C ) Colony formation (B) and size (C, mm 2 ) of Translator SLAM-LKS sorted on the basis of IRES/Cap ( n = 6). ( D and E ) Colony formation (D) and size (E, mm 2 ) of Translator GMP sorted on the basis of IRES/Cap ( n = 9). ( F ) Total protein synthesis rates of Translator GMP based on IRES/Cap measured by AF647-OPP MFI ( n = 3). ( G ) Colony formation of EMCV-IRES reporter-transduced CD34+CD90+EPCR+CD45RA− human cord blood sorted on the basis of IRES/Cap ( n = 3). ( H and I ) Colony formation of SLAM-LKS (H) and GMPs (I) based on RUNX1-IRES/Cap sorted from transplant recipients ( n = 3 and 2, respectively). ( J ) Primary competitive transplant schema. Twenty CD45.2 SLAM-LKS sorted on the basis of IRES/Cap versus 200,000 CD45.1 whole BM cells in lethally irradiated CD45.1 recipients. Peripheral blood chimerism analyzed every four weeks through week 16. Created in BioRender. Li, D. (2026) https://BioRender.com/d7k2abt . ( K ) Total chimerism Translator SLAM-LKS based on IRES/Cap in primary recipients evaluated through week 16 ( n = 12). ( L ) Secondary competitive transplant schema. One million total BM cells were collected from primary recipients (16 weeks posttransplantation) and transplanted to lethally irradiated CD45.1 recipients. Created in BioRender. Li, D. (2026) https://BioRender.com/d7k2abt . ( M ) Total chimerism Translator SLAM-LKS based on IRES/Cap in secondary recipients through week 16 ( n = 12). ( N and O ) Representative image of the culture of a single low (N) or high (O) IRES/Cap SLAM-LKS after 5 days of culture. ( P and Q ) Pie charts depicting the percentage of wells with megakaryocytes (MK) after 5 days of culturing a single low (P) or high (Q) IRES/Cap Translator SLAM-LKS ( n = 20). Data show individual replicates and means ± SEM. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001. Significance was assessed using a two-way ANOVA with Dunnett’s one-way ANOVA (B to I) or an ANOVA with Tukey’s multiple comparisons test (K and M).

Article Snippet: Heterozygous transgene positive founders were bred with WT CD45.2 C57BL/6J mice purchased from the Jackson Laboratory.

Techniques: Irradiation

Journal: Cancer Research

Article Title: Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression

doi: 10.1158/0008-5472.CAN-25-3092

Figure Lengend Snippet: ABHD16A promotes ILC3 aggregation and IL22 release in the immune microenvironment. A, A total of 5 × 10 6 MFC-luc or Abhd16a -knockdown MFC-luc cells were injected into the epidermis of the greater curvature of the stomach in the 615 mice. Tumor growth was monitored weekly through bioluminescence imaging from the first day of MFC cell inoculation. Scale bar, 2.000e+4 – 5.000e + 5 p/s/cm 2 /sr. B and C, Representative images of the excised stomachs ( B ) and the orthotopic tumor volume ( C ) on day 28 after gastric cancer cells were implanted orthotopically ( n = 9 per group). D, H&E images of the stomach, peritoneum, liver, intestine, and lung on day 42 after MFC or Abhd16a -knockdown MFC cells were implanted into 615 mice. Scale bar, 400 μm. E–G, Representative images ( E ), volume ( F ), and weight ( G ) of control and Abhd16a -knockdown subcutaneous tumors ( n = 5 per group). H, IHC images of PD-L1 in control and Abhd16a -knockdown orthotopic and subcutaneous gastric cancer tissues. Scale bar, 500 μm. I, Detection of mouse cytokines in the TIF of orthotopic gastric cancer tissues by proteome profiler mouse XL cytokine array. J, Flow cytometry gating strategy and frequencies of ILC3s and Th17/Th22 cells out of CD45 + cells in control and Abhd16a -knockdown gastric cancer tissues in 615 mice. K, Flow cytometry gating strategy and frequencies of ILC3s out of ILCs in control and Abhd16a -knockdown gastric cancer tissues in Rag1 −/− mice. ***, P < 0.001.

Article Snippet: FITC anti-human CD3 (Thermo Fisher Scientific, cat. #11-0038-42, RRID: AB_2043831, 5 μL/1 × 10 6 cells), PE anti-human CD127 (BioLegend, cat. #351304, RRID: AB_10720185, 5 μL/1 × 10 6 cells), Brilliant Violet 421 anti-human CD294 (BioLegend, cat. #350112, RRID: AB_2562468, 5 μL/1 × 10 6 cells), APC anti-human CD117 (BioLegend, cat. #313206, RRID: AB_314985, 5 μL/1 × 10 6 cells), FITC anti-human CD19 (eBioscience, cat. #11-0199-42, RRID: AB_10669461, 5 μL/1 × 10 6 cells), FITC anti-human CD14 (eBioscience, cat. #11-0149-42, RRID: AB_10597597, 5 μL/1 × 10 6 cells), BV650 anti-human CD45 (eBioscience, cat. #416-0459-42, RRID: AB_2925684, 5 μL/1 × 10 6 cells), iFluor 647 anti-Ki67 (HUABIO, cat. #HA720163F, RRID: AB_3072100, 1 μL/1 × 10 6 cells), PE/Cyanine7 anti-human CD274 (Elabscience, cat. #E-AB-F1133H, 5 μL/1 × 10 6 cells), and PE anti-human CD273 (Elabscience, cat. #E-AB-F1175D, 5 μL/1 × 10 6 cells).

Techniques: Knockdown, Injection, Imaging, Control, Flow Cytometry

Journal: Cancer Research

Article Title: Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression

doi: 10.1158/0008-5472.CAN-25-3092

Figure Lengend Snippet: High expression of ABHD16A mediates the cumulative release of LysoPS into the TME. A, mIF images of the orthotopic gastric cancer (GC) tissues stained for PD-L1, RORC, and CD3. Yellow, PD-L1 + tumor cells; red, RORC + ILC3s; green, CD3 + cells. Scale bars, 100 μm (left) and 25 μm (right). B and C, Flow cytometry gating strategy and frequencies of ILC3s ( B ) and proportions of ILC3s in total CD45 + cells ( C ) isolated from control and Abhd16a -knockdown orthotopic and subcutaneous gastric cancer tissues ( n = 3 per group). D and E, Flow cytometry gating strategy and frequencies of ILC3s ( D ) and proportions of ILC3s in total CD45 + cells ( E ) derived from the peripheral blood of healthy controls (HC) and patients with gastric cancer ( n = 8 per group). F and G, LC-MS/MS analysis of 18:0 and 18:1 LysoPS in the control and Abhd16a -knockdown orthotopic ( F ) and subcutaneous ( G ) gastric cancer tissues ( n = 3 per group). H–J, ELISA was used to measure levels of LysoPS in gastric cancer cell supernatant ( H ), TIF ( I ), and in vitro tumor culture supernatant ( J ) of control and Abhd16a -knockdown orthotopic and subcutaneous gastric cancer tumors ( n = 3 per group). K, LysoPS levels in the serum of healthy controls and patients with gastric cancer detected by ELISA. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: FITC anti-human CD3 (Thermo Fisher Scientific, cat. #11-0038-42, RRID: AB_2043831, 5 μL/1 × 10 6 cells), PE anti-human CD127 (BioLegend, cat. #351304, RRID: AB_10720185, 5 μL/1 × 10 6 cells), Brilliant Violet 421 anti-human CD294 (BioLegend, cat. #350112, RRID: AB_2562468, 5 μL/1 × 10 6 cells), APC anti-human CD117 (BioLegend, cat. #313206, RRID: AB_314985, 5 μL/1 × 10 6 cells), FITC anti-human CD19 (eBioscience, cat. #11-0199-42, RRID: AB_10669461, 5 μL/1 × 10 6 cells), FITC anti-human CD14 (eBioscience, cat. #11-0149-42, RRID: AB_10597597, 5 μL/1 × 10 6 cells), BV650 anti-human CD45 (eBioscience, cat. #416-0459-42, RRID: AB_2925684, 5 μL/1 × 10 6 cells), iFluor 647 anti-Ki67 (HUABIO, cat. #HA720163F, RRID: AB_3072100, 1 μL/1 × 10 6 cells), PE/Cyanine7 anti-human CD274 (Elabscience, cat. #E-AB-F1133H, 5 μL/1 × 10 6 cells), and PE anti-human CD273 (Elabscience, cat. #E-AB-F1175D, 5 μL/1 × 10 6 cells).

Techniques: Expressing, Staining, Flow Cytometry, Isolation, Control, Knockdown, Derivative Assay, Liquid Chromatography with Mass Spectroscopy, Enzyme-linked Immunosorbent Assay, In Vitro

Journal: Cancer Research

Article Title: Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression

doi: 10.1158/0008-5472.CAN-25-3092

Figure Lengend Snippet: LysoPS enhances the proliferation and activation of ILC3s in the TME. A, Flow cytometry was performed to detect the frequencies of Ki67 + MNK3 under treatment with LysoPS (10 μmol/L) for 24 hours. B and C, Flow cytometry gating strategy and frequencies of ILC3s ( B ) and proportions of ILC3s in total CD45 + cells ( C ) in the isolated human PBMCs treated with exogenous LysoPS (10 μmol/L) for 24 hours ( n = 4 per group). D, Numbers of ILC3s treated with LysoPS. Cell counts were recorded daily for 5 days ( n = 3 per group). E, ELISA analysis of IL22 in the supernatant of MNK3 cells stimulated with incremental concentrations (0, 5, 10, and 20 μmol/L) of LysoPS ( n = 3 per group). F, Frequencies of IL22 + ILC3s out of total ILC3s isolated from control and Abhd16a -knockdown orthotopic and subcutaneous gastric cancer tissues ( n = 3 per group). G and H, ELISA analysis of IL22 in the TIF of control and Abhd16a -knockdown orthotopic and subcutaneous gastric cancer tissues ( G ) and IL22 in the serum of healthy controls (HC) and patients with gastric cancer ( H ). I, Correlation analysis between IL22 and LysoPS in the serum of patients with gastric cancer. J, The levels of LysoPS and IL22 in the serum of patients with PNI − and PIN + gastric cancer measured by ELISA. K, ROC analysis of the diagnostic value of LysoPS and IL22 for gastric cancer. **, P < 0.01; ***, P < 0.001.

Article Snippet: FITC anti-human CD3 (Thermo Fisher Scientific, cat. #11-0038-42, RRID: AB_2043831, 5 μL/1 × 10 6 cells), PE anti-human CD127 (BioLegend, cat. #351304, RRID: AB_10720185, 5 μL/1 × 10 6 cells), Brilliant Violet 421 anti-human CD294 (BioLegend, cat. #350112, RRID: AB_2562468, 5 μL/1 × 10 6 cells), APC anti-human CD117 (BioLegend, cat. #313206, RRID: AB_314985, 5 μL/1 × 10 6 cells), FITC anti-human CD19 (eBioscience, cat. #11-0199-42, RRID: AB_10669461, 5 μL/1 × 10 6 cells), FITC anti-human CD14 (eBioscience, cat. #11-0149-42, RRID: AB_10597597, 5 μL/1 × 10 6 cells), BV650 anti-human CD45 (eBioscience, cat. #416-0459-42, RRID: AB_2925684, 5 μL/1 × 10 6 cells), iFluor 647 anti-Ki67 (HUABIO, cat. #HA720163F, RRID: AB_3072100, 1 μL/1 × 10 6 cells), PE/Cyanine7 anti-human CD274 (Elabscience, cat. #E-AB-F1133H, 5 μL/1 × 10 6 cells), and PE anti-human CD273 (Elabscience, cat. #E-AB-F1175D, 5 μL/1 × 10 6 cells).

Techniques: Activation Assay, Flow Cytometry, Isolation, Enzyme-linked Immunosorbent Assay, Control, Knockdown, Diagnostic Assay

Journal: Cancer Research

Article Title: Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression

doi: 10.1158/0008-5472.CAN-25-3092

Figure Lengend Snippet: ABHD16A–LysoPS enhances ILC3 activation via the GPR34–STAT3–AKT signaling pathway. A, MNK3 cells were treated with or without LysoPS (10 μmol/L); the levels of IL22, pAKT, AKT, pSTAT3, and STAT3 were assessed by Western blotting. B, Western blotting was performed to check IL22, pAKT, AKT, pSTAT3, and STAT3 levels in MNK3 cells cocultured with MFC cells, Abhd16a -knockdown MFC cells, or Abhd16a -knockdown MFC cells with simultaneous supplementation of LysoPS (10 μmol/L). C, Western blotting was conducted to assess IL22, pAKT, AKT, pSTAT3, and STAT3 levels in MNK3, Gpr34 -knockdown MNK3, and perifosine-pretreated MNK3 cells, all under LysoPS supplementation (10 μmol/L). D, ELISA analysis of IL22 production in supernatants from MNK3 cells under the same treatments as in B . E, ELISA analysis of IL22 production in the supernatant from MNK3 cells under the same treatments as in C . F, ELISA analysis of IL22 production by ILC3s and ILC3s treated with LysoPS in the presence or absence of perifosine. G and H, LysoPS (2.5 mg/kg) was administered to Abhd16a -knockdown gastric cancer mice and control mice in the presence or absence of a GPR34 inhibitor (GPR34 receptor antagonist 2, 20 mg/kg) or AKT inhibitor (perifosine, 20 mg/kg). The representative living images on days 3, 10, and 17 ( G ) and tumor volume at the end of the experiment ( H ) are shown. Scale bar, 1.000e+4 – 1.000e + 5 p/s/cm 2 /sr; n = 5 per group. I–K, The proportions of ILC3s ( I ), pSTAT3 + ILC3s ( J ), and IL22 + ILC3s ( K ) in total CD45 + cells derived from orthotopic gastric cancer tissues. Mice treatments were the same as G and H . **, P < 0.01; ns, nonsignificant.

Article Snippet: FITC anti-human CD3 (Thermo Fisher Scientific, cat. #11-0038-42, RRID: AB_2043831, 5 μL/1 × 10 6 cells), PE anti-human CD127 (BioLegend, cat. #351304, RRID: AB_10720185, 5 μL/1 × 10 6 cells), Brilliant Violet 421 anti-human CD294 (BioLegend, cat. #350112, RRID: AB_2562468, 5 μL/1 × 10 6 cells), APC anti-human CD117 (BioLegend, cat. #313206, RRID: AB_314985, 5 μL/1 × 10 6 cells), FITC anti-human CD19 (eBioscience, cat. #11-0199-42, RRID: AB_10669461, 5 μL/1 × 10 6 cells), FITC anti-human CD14 (eBioscience, cat. #11-0149-42, RRID: AB_10597597, 5 μL/1 × 10 6 cells), BV650 anti-human CD45 (eBioscience, cat. #416-0459-42, RRID: AB_2925684, 5 μL/1 × 10 6 cells), iFluor 647 anti-Ki67 (HUABIO, cat. #HA720163F, RRID: AB_3072100, 1 μL/1 × 10 6 cells), PE/Cyanine7 anti-human CD274 (Elabscience, cat. #E-AB-F1133H, 5 μL/1 × 10 6 cells), and PE anti-human CD273 (Elabscience, cat. #E-AB-F1175D, 5 μL/1 × 10 6 cells).

Techniques: Activation Assay, Western Blot, Knockdown, Enzyme-linked Immunosorbent Assay, Control, Derivative Assay