Journal: Gut microbes

Article Title: Systemic exposure to bacterial amyloid curli alters the gut mucosal immune response and the microbiome, exacerbating Salmonella -induced arthritis.

doi: 10.1080/19490976.2023.2221813

Figure Lengend Snippet: Figure 5. Expression of chemokines in the cecum and colon. Levels of Cxcl5, Ccl3, and Cxcl1 determined by RT-qPCR in RNA extracted 24 hours after indicated treatment from (a) colon and (b) cecum of C57BL/6 mice from either Jackson Labs or Taconic Farms. Data were normalized to data from Jackson Labs mice treated with PBS. Each data point represents one mouse sample. Mean and standard error were calculated by averaging results from two independent experiments. Significance was determined using a Mann-Whitney test; **p < 0.01, ***p < 0.001.

Article Snippet: Levels of Cxcl5, Ccl3, and Cxcl1 determined by RT-qPCR in RNA extracted 24 hours after indicated treatment from (a) colon and (b) cecum of C57BL/6 mice from either Jackson Labs or Taconic Farms.

Techniques: Expressing, Quantitative RT-PCR, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: Cancer-derived exosomal HSPC111 promotes colorectal cancer liver metastasis by reprogramming lipid metabolism in cancer-associated fibroblasts

doi: 10.1038/s41419-022-04506-4

Figure Lengend Snippet: Overlapping of genes resulting from comparison of downregulated genes in LX-2 cells incubated with Exo HCT116 KD and upregulated genes in LX-2 cells incubated with Exo SW480 OE ( A ). Heat map of transcriptome alterations after incubating LX-2 cells with Exo HCT116 KD and Exo HCT116 KD Ctrl ( B ). Heat map of transcriptome alterations after incubating LX-2 cells with Exo SW480 OE and Exo SW480 OE Ctrl ( C ). The relative mRNA level of CXCL5 was analyzed in a group of independent samples ( D ). Western blot showed protein level of CXCL5 in LX-2 cells incubated with Exo HCT116 KD ( E ) and Exo SW480 OE (F) and their relative control respectively. ELISA analysis of CXCL5 level in culture medium from LX-2 cells incubated with Exo HCT116 KD ( G ) and Exo SW480 OE ( H ) and their relative control respectively. Immunofluorescence staining of α-SMA, FAP and CXCL5 in LX-2 cells incubated with Exo HCT116 KD ( I ) and Exo SW480 OE ( J ) and their relative control respectively. Scale bar = 20 μm. Each experiment was performed in triplicate. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, ns not significant.

Article Snippet: Cells were cultured in the presence of appropriate exosomes (20 μg/mL) for 48 h before supernatants were collected, and the CXCL5 level was detected by CXCL5 ELISA kit (EK0728, BOSTER) according to the manufacturer’s protocols.

Techniques: Comparison, Incubation, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining

Journal: Cell Death & Disease

Article Title: Cancer-derived exosomal HSPC111 promotes colorectal cancer liver metastasis by reprogramming lipid metabolism in cancer-associated fibroblasts

doi: 10.1038/s41419-022-04506-4

Figure Lengend Snippet: The effects of acetate treatment on acetyl-CoA levels in LX-2 cells incubated with Exo HCT116 ( A ) and Exo SW480 ( B ). Western blot showed the effects of acetate treatment on CXCL5 and H3K27ac levels in LX-2 cells incubated with Exo HCT116 ( C ) and Exo SW480 ( D ). E The effects of Trichostatin A (TSA) treatment on CXCL5 and H3K27ac levels in LX-2 cells incubated with Exo HCT116 KD and their relative control. F The effects of C646 treatment on CXCL5 and H3K27ac levels in LX-2 cells incubated with Exo SW480 OE and their relative control. G , H Chromatin immunoprecipitation (ChIP) assays using IgG as a control were performed with antibody against H3K23ac. Examination of H3K27 acetylation status in CXCL5 gene promoter region in LX-2 cells incubated with Exo HCT116 KD and Exo HCT116 KD Ctrl or in LX-2 cells incubated with Exo HCT116 KD and Exo HCT116 KD Ctrl and treated with TSA ( G ). Examination of H3K27 acetylation status in CXCL5 gene promoter region in LX-2 cells incubated with Exo SW480 OE and Exo SW480 OE Ctrl or in LX-2 cells incubated with Exo SW480 OE and Exo SW480 OE Ctrl and treated with C646 ( H ). Each experiment was performed in triplicate. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were cultured in the presence of appropriate exosomes (20 μg/mL) for 48 h before supernatants were collected, and the CXCL5 level was detected by CXCL5 ELISA kit (EK0728, BOSTER) according to the manufacturer’s protocols.

Techniques: Incubation, Western Blot, Control, Chromatin Immunoprecipitation

Journal: Cell Death & Disease

Article Title: Cancer-derived exosomal HSPC111 promotes colorectal cancer liver metastasis by reprogramming lipid metabolism in cancer-associated fibroblasts

doi: 10.1038/s41419-022-04506-4

Figure Lengend Snippet: Western blot showed the protein levels of HSPC111, CXCR2 and EMT-relative genes in CAFs CM-incubated HCT116 ( A ) and SW480 cells ( B ). Migration of HCT116 ( C ) and SW480 cells ( D ) were analyzed by transwell assay after CAFs CM treatment. Scale bar = 200 μm. Western blot showed the protein levels of HSPC111, CXCR2 and EMT-relative genes in CAFs CM- and CXCL5 neutralizing antibody-incubated HCT116 ( E ) and SW480 cells ( F ). G Migration of HCT116 cells were analyzed by wound-healing assay after CAFs CM and CXCL5 neutralizing antibody treatment. Scale bar = 100 μm. Western blot showed the protein levels of HSPC111, CXCR2 and EMT-relative genes in CAFs CM- and CXCR2 inhibitor (navarixin)-incubated HCT116 ( H ) and SW480 cells ( I ). J Migration of HCT116 cells were analyzed by wound-healing assay after CAFs CM and CXCR2 inhibitor (navarixin) treatment. Scale bar = 100 μm. K IVIS imaging on experimental liver metastasis of indicated mice treated with Exo HCT116 KD or their relative control and with/without navarixin. Luciferase-labeled HCT116 cells were used to perform experimental liver metastasis model ( n = 3). L Representative pictures and quantitative results of HE staining of liver tissue sections from indicated mice. Scale bar = 500 μm. Each experiment was performed in triplicate. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were cultured in the presence of appropriate exosomes (20 μg/mL) for 48 h before supernatants were collected, and the CXCL5 level was detected by CXCL5 ELISA kit (EK0728, BOSTER) according to the manufacturer’s protocols.

Techniques: Western Blot, Incubation, Migration, Transwell Assay, Wound Healing Assay, Imaging, Control, Luciferase, Labeling, Staining

Journal: Cell Death & Disease

Article Title: Cancer-derived exosomal HSPC111 promotes colorectal cancer liver metastasis by reprogramming lipid metabolism in cancer-associated fibroblasts

doi: 10.1038/s41419-022-04506-4

Figure Lengend Snippet: Exosomal HSPC111 derived from CRC cells phosphorylates ACLY in CAFs, leading to the increased levels of acetyl-CoA and histone acetylation to secrete CXCL5, and resulting in CRC cells colonized in liver via the CXCL5-CXCR2 axis.

Article Snippet: Cells were cultured in the presence of appropriate exosomes (20 μg/mL) for 48 h before supernatants were collected, and the CXCL5 level was detected by CXCL5 ELISA kit (EK0728, BOSTER) according to the manufacturer’s protocols.

Techniques: Derivative Assay