Journal: Advanced Science
Article Title: The Characteristics of Tumor Microenvironment by the Single Cell Profile in Endometrial Cancer of no Specific Molecular Profile (NSMP) Subtype
doi: 10.1002/advs.78332
Figure Lengend Snippet: SPP1+ macrophages promote T cell stress and maintain epithelial cell stemness. (A, B) Western blot and RT‐qPCR results of SPP1 expression in macrophages with SPP1 knockdown. (C, D) Western blot and RT‐qPCR results of SPP1 expression in macrophages with overexpression of SPP1. (E) Western blot analysis of the protein expression of HSP70, LAG3, and PD‐1 in Jurkat cells after co‐culture with control and SPP1 knockdown macrophages. (F) qRT‐PCR analysis of the transcriptional level of HSPA1A and HSPA1B in Jurkat cells after co‐culture with control and SPP1 knockdown macrophages. (G) Western blot analysis of the protein expression of HSP70, LAG3, and PD‐1 in Jurkat cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. (H) qRT‐PCR analysis of the transcriptional level of HSPA1A and HSPA1B in Jurkat cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. (I) Western blot analysis of the protein expression of CD44 and CD133 in Ishikawa cells after co‐culture with control and SPP1 knockdown macrophages. (J) Western blot analysis of the protein expression of CD44 and CD133 in Ishikawa cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. (K) Sphere formation assay was used to detect the stemness ability of Ishikawa cells after co‐culture with control and SPP1 knockdown macrophages. (L) A sphere formation assay was used to detect the stemness ability of Ishikawa cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant.
Article Snippet: The primary antibodies were: EPCAM (21050‐1‐AP, Proteintech), CD68 (25747‐1‐AP, Proteintech), SPP1 (22952‐1‐AP, Proteintech), CD4 (67786‐1‐lg, Proteintech) and CD44 (15675‐1‐AP, Proteintech).
Techniques: Western Blot, Quantitative RT-PCR, Expressing, Knockdown, Over Expression, Analysis, Co-culture Assay, Control, Tube Formation Assay