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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.

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

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.

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