Journal: Signal Transduction and Targeted Therapy

Article Title: XBP1 regulates the protumoral function of tumor-associated macrophages in human colorectal cancer

doi: 10.1038/s41392-021-00761-7

Figure Lengend Snippet: Effect of XBP1 activation on the pro-tumor function of TAMs. a Western blot analysis of XBP1s expression in indicated macrophages. b Representative micrograph showing tumor formation in NOD/SCID mice injected subcutaneously (s.c.) with luciferase tumor cells (CT26-luciferase) and two groups of macrophages: CT26 + BMDMs (black arrows) and CT26 + miTAMs (magenta arrows). c Growth curves of the two groups in b . d Western blot analysis of XBP1 expression in sgCon or sgXbp1 TAMs. e Representative photograph showing tumor formation in NOD/SCID mice injected s.c. with luciferase tumor cells (CT26-luciferase) and two groups of miTAMs: CT26 + sgCon miTAMs (black arrows); and CT26 + sgXbp1 miTAMs (blue arrows). f Growth curves of the two groups in e . **** P < 0.0001; Repeated measurement and analysis. g CT26 cells, mixed with: Con TAMs; Xbp1s TAMs; sgCon TAMs; and sgXbp1 TAMs, were orthotopically injected into the wall of the cecum ( n = 5 mice per group). Macroscopic appearance of the CRC orthotopic tumors with each indicated treatment. Black arrows indicate macroscopic polyps. Scale bar, 1 cm. h Representative HE staining of liver metastasis in mice xenografted of the four groups in g . Scale bar, 100 μm. i , j Statistical analysis of the orthotopic CRC tumor weight ( i ) and the clone number of liver metastasis ( j ). * P < 0.05, *** P < 0.001; t -test. k Incidence of orthotopic CRC tumor formation and liver metastasis analysis. l Representative images of CT26 pulmonary metastases induced by tail vein injection in NOD/SCID. CT26 cells were mixed with: Con miTAMs; Xbp1s miTAMs; sgCon miTAMs; and sgXbp1 miTAMs. HE staining demonstrating the histology of tumors formed in the lungs; scale bar, 500 μm. m Pulmonary metastatic nodule numbers in i . ( n = 4 mice per group); ** P < 0.01, *** P < 0.001; t -test

Article Snippet: Ana-1 (EP-CL-0023, RRID:CVCL_0142) and CT26 cells (EP-CL-0071, RRID:CVCL_7254) were from Elabscience (Texas, USA).

Techniques: Activation Assay, Western Blot, Expressing, Injection, Luciferase, Staining

Journal: Signal Transduction and Targeted Therapy

Article Title: XBP1 regulates the protumoral function of tumor-associated macrophages in human colorectal cancer

doi: 10.1038/s41392-021-00761-7

Figure Lengend Snippet: Effect of XBP1 on macrophages phagocytosis. a Representative images of phagocytosis assays using RFP-labeled human CRC cell, DLD1cells (DLD1-RFP) and sgCon or sgXBP1 hiTAMs ( n = 3). Yellow arrows denote phagocytic events. Scale bar = 200 μm. b Representative images of phagocytosis assays using RFP-labeled mouse CT26 cells (CT26-RFP) and sgCon or sgXbp1 miTAMs ( n = 3). Yellow arrows denote phagocytic events. Scale bar = 200 μm. c Results of phagocytosis assays of the two groups in a , b . ** P < 0.01; t -test. d Relative mRNA levels of XBP1 and phagocytosis-associated genes in BMDM, sgCon miTAMs and sgXbp1 miTAMs, validated by RT-qPCR. Bars represent mean ± SD of three experimental replicates. * P < 0.05, ** P < 0.01, *** P < 0.001; t -test. e Expression of THBS1 and SIRPα versus XBP1 in TAMs sorted from CRC patients ( n = 27 total). r , Spearman’s rank correlation test. f Correlation of XBP1 with THBS1 and SIRPα in CRC patients. The association was analyzed using coefficient measures of the linear relationships in the public GEO database (GSE68468). g Track view of Erdj4 , Thbs1 , and Sipra ChIP-seq density upon silencing of Xbp1 in the ChIP-seq online database (GSE86048). h ChIP-qPCR experiments measuring XBP1 binding on Erdj4 , Thbs1 , and Sipra segments. Bars represent mean ± SD of three experimental replicates. * P < 0.05, ** P < 0.01. P -values were determined using t -test

Article Snippet: Ana-1 (EP-CL-0023, RRID:CVCL_0142) and CT26 cells (EP-CL-0071, RRID:CVCL_7254) were from Elabscience (Texas, USA).

Techniques: Labeling, Quantitative RT-PCR, Expressing, ChIP-sequencing, ChIP-qPCR, Binding Assay