Journal: Frontiers in Immunology

Article Title: UPF1-dependent glycolysis driven by LINC00887 facilitates macrophage M2 polarization in clear cell renal cell carcinoma

doi: 10.3389/fimmu.2026.1833383

Figure Lengend Snippet: Overexpression of UPF1 in ccRCC inhibits M2 macrophage polarization and enhances the tumor suppressor function of CD8 + T cells, Caki-1 cell groups of sh-NC+PBS, sh-UPF1+PBS, or sh-UPF1 + 2-DG, and the conditioned medium was used to culture and polarize M0 macrophages. The polarized macrophages were cultured with activated CD8 + T cells. (A) CFSE assessed CD8 + T cell proliferative capacity. (B) Flow cytometry measured apoptosis rate of CD8 + T cells in different co-culture systems. (C) CD8 + T cells from culture systems were co-cultured with normal Caki-1 cells, with LDH assays determining cytotoxic effect of CD8 + T cells. *, p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; 2-DG, 2-Deoxy-D-glucose; CFSE, Carboxyfluorescein succinimidyl ester.

Article Snippet: Antibodies included: FITC Mouse monoclonal to CD68 (ab134351, abcam, UK), Pacific Blue anti-human CD86 Antibody (305417, yajimall, China), CD206 (MMR) Monoclonal Antibody (19.2)-PE (12-2069-42, ThermoFisher, USA), PerCP/Cyanine5.5 Anti-Mouse F4/80 Antibody (E-AB-F0995J, Elabscience, China), PE Anti-CD206 mouse antibody (E-AB-F1135D, Elabscience, China), APC Anti-mouse CD3 antibody (E-AB-F1013E, Elabscience, China), PE Anti-mouse CD8 alpha antibody (E-AB-F1104D, Elabscience, China).

Techniques: Over Expression, Cell Culture, Flow Cytometry, Co-culture Assay

Journal: Frontiers in Immunology

Article Title: UPF1-dependent glycolysis driven by LINC00887 facilitates macrophage M2 polarization in clear cell renal cell carcinoma

doi: 10.3389/fimmu.2026.1833383

Figure Lengend Snippet: LINC00887/UPF1-mediated aerobic glycolysis leads to lactic acid accumulation and facilitates immune evasion in ccRCC in vivo , A syngeneic tumor model was developed by injecting BALB/c mice with Renca cells expressing sh-NC or sh-LINC00887, followed by treatment with sh-NC or sh-UPF1 lentivirus. A-C: Assessment of tumor size (A) , volume (B) , and weight (C) . (D, E) qRT-PCR analyses for LINC00887 (D) and UPF1 (E) mRNA levels. (F) IHC analysis for UPF1, LDH-A, KI67 protein levels. (G) Kit assay for lactate levels in tumors. (H) Proportion of M2 macrophages (F4/80+CD206+) by flow cytometry. (I) Immunofluorescence assay for CD206+ macrophages. (J) Proportion of CD8 + T cells (CD3 + CD8 + ) in tumor tissues. ns, no significant difference. * p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; LDH-A, Lactate dehydrogenase A; CD206, Mannose receptor C-type 1; CD3, Cluster of differentiation 3; CD8, Cluster of differentiation 8.

Article Snippet: Antibodies included: FITC Mouse monoclonal to CD68 (ab134351, abcam, UK), Pacific Blue anti-human CD86 Antibody (305417, yajimall, China), CD206 (MMR) Monoclonal Antibody (19.2)-PE (12-2069-42, ThermoFisher, USA), PerCP/Cyanine5.5 Anti-Mouse F4/80 Antibody (E-AB-F0995J, Elabscience, China), PE Anti-CD206 mouse antibody (E-AB-F1135D, Elabscience, China), APC Anti-mouse CD3 antibody (E-AB-F1013E, Elabscience, China), PE Anti-mouse CD8 alpha antibody (E-AB-F1104D, Elabscience, China).

Techniques: In Vivo, Expressing, Quantitative RT-PCR, Analysis, Flow Cytometry, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: Protective effects of YCHD on the autoimmune hepatitis mice model induced by Ad-CYP2D6 through modulating the Th1/Treg ratio and intestinal flora

doi: 10.3389/fimmu.2024.1488125

Figure Lengend Snippet: Effect of YCHD on Th1 and Treg cell differentiation. Lymphocytes isolated from the spleen and hepatic lymph nodes of the C57BL/6 mice were stimulated with or without PMA (50ng/ml), ionomycin (1μg/ml), and BFA (10μg/ml) for 5h at 37°. (A, B) The gating strategy of Th1 ang Treg cells isolated from the spleen defined as CD3 + CD4 + IFN-γ + cells and CD3 + CD4 + CD25 + FOXP3 + cells and the representative dot plots from each group are shown. (C) Frequency Th1 and Treg cells in the spleen of mice (n=8) in each group were analyzed by flow cytometry. (D, E) The Th1 and Treg population in the hepatic hilar lymph nodes was analyzed by staining the cell surface CD4 antigen and IFN-γ antigen, as well as by staining of cell surface CD4 antigen, CD25 antigen, and intracellular Foxp3, in the CD3 + T cells population. (F) Frequency Th1 and Treg cells in the hepatic hilar lymph nodes of mice (n=8) in each group were also analyzed by flow cytometry. *P<0.05, **P<0.01, compared with the control group. ##P<0.01, compared with the untreated group.

Article Snippet: Cells were blocked with 50μl blocking buffer (2% goat serum, 2% fetal bovine serum, and 96% PBS) for 30 minutes at 4 °, 5μl PerCP Anti-Mouse CD3 Antibody (E-AB-F1013F, Elabscience) and 0.5μl FITC anti-mouse CD4 (B374032, BioLegend) were added for surface staining (incubated at 4°C for 40 minutes).

Techniques: Cell Differentiation, Isolation, Flow Cytometry, Staining, Control