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: UPF1 regulates aerobic glycolysis in ccRCC to inhibit macrophage M2 polarization, 786-O cell groups of sh-NC+PBS, sh-UPF1+PBS, or sh-UPF1 + 2-DG. (A) Immunoblotting detected protein levels of UPF1, LDH-A, HK2, and GLUT1 in each group of cells. (B) Seahorse Extracellular Flux Analyzer tested ECAR of each group, calculating glycolytic rate, glycolytic capacity, and glycolytic reserve. (C) Seahorse Extracellular Flux Analyzer tested OCR, calculating basal respiration, ATP production, and maximum respiration. (D–F) Assay kits for glucose consumption (D) , lactate production (E) , and ATP production (F) . Conditioned medium from differently treated Caki-1 cells was used to culture M0 macrophages, (G, H) Flow cytometry quantification of M1 macrophages (CD68 + CD86 + ) and M2 macrophages (CD68 + CD206 + ). (I, J) qRT-PCR analysis of M1 markers [TNF-α, iNOS, IL-1β; (I) ] and M2 markers [Arg1, Fizz1, IL-10; (J) ]. *, p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; 2-DG, 2-Deoxy-D-glucose; LDH-A, Lactate dehydrogenase A; HK2, Hexokinase 2; GLUT1, Glucose transporter 1; ECAR, Extracellular acidification rate; OCR, Oxygen consumption rate; ATP, Adenosine triphosphate; CD68, Cluster of differentiation 68; CD86, Cluster of differentiation 86; CD206, Mannose receptor C-type 1; TNF-α, Tumor necrosis factor-alpha; iNOS, Inducible nitric oxide synthase; IL-1β, Interleukin - 1β; Arg1, Arginase 1; Fizz1, Found in inflammatory zone 1; IL-10, Interleukin -10.

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: Western Blot, Flow Cytometry, Quantitative RT-PCR, Analysis

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