Journal: Cell Communication and Signaling : CCS

Article Title: The tumor promoter cysteinyl leukotriene receptor 1 regulates PD-L1 expression in colon cancer cells via the Wnt/β-catenin signaling axis

doi: 10.1186/s12964-023-01157-6

Figure Lengend Snippet: List and details of reagents and antibodies used in this study

Article Snippet: Total β-Catenin (D10A8) XP® Rabbit mAb , BDBioscience , #8480 , 1:5000 , Immunoblotting.

Techniques: Dilution Assay, Western Blot, Recombinant, Fluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: The tumor promoter cysteinyl leukotriene receptor 1 regulates PD-L1 expression in colon cancer cells via the Wnt/β-catenin signaling axis

doi: 10.1186/s12964-023-01157-6

Figure Lengend Snippet: Antagonizing CysLT 1 R affects PD-L1 expression in different mouse xenograft models. Immunofluorescence images showing the expression of β-catenin (green) and PD-L1 (red) in A HT-29 and B HCT116 xenografts in mice treated with DMSO or Montelukast (Mo). Representative graphs showing the MFI (Mean fluorescence intensity) of PD-L1 ( n = 4 mice per group in each xenograft condition). The scale bars are indicated in the images. The white dotted line marks the border of the xenograft section. Western blots of C HT-29 cell xenografts and D HCT116 cell xenografts. The blots show the expression of the indicated proteins of interest. Representative densitometric analysis results are shown in the graphs for HT-29 and HCT116 cells. In all the western blot panels, GAPDH served as the loading control. MW, relative molecular weight expressed in kilodaltons (kDa). The MFI in all confocal images was calculated with ImageJ software (NIH, USA). Mean ± SEM, * P < 0.05, *** P < 0.001, two-tailed unpaired t test

Article Snippet: Total β-Catenin (D10A8) XP® Rabbit mAb , BDBioscience , #8480 , 1:5000 , Immunoblotting.

Techniques: Expressing, Immunofluorescence, Fluorescence, Western Blot, Control, Molecular Weight, Software, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: The tumor promoter cysteinyl leukotriene receptor 1 regulates PD-L1 expression in colon cancer cells via the Wnt/β-catenin signaling axis

doi: 10.1186/s12964-023-01157-6

Figure Lengend Snippet: The CysLT 1 R-specific antagonist Mo shows combinatorial effects with an anti-PD-L1 neutralizing antibody. A Western blots showing the expression of the indicated proteins in RKO CC cells treated with Montelukast (Mo, 10 µM) or Atezolizumab (20 ng/mL) alone or together. The blots are representative of three replicates, and the results are shown in the densitometry graphs. B Immunofluorescence images showing PD-L1 expression in RKO cells. Cells were stimulated with Mo (10 µM) for 30 min or Atezolizumab (20 ng/mL) for 24 h alone or in combination with pretreatment with Mo (10 µM, 30 min) followed by Atezolizumab ( 20 ng/mL) for 24 h. The left panel shows the merged confocal images in each group with nuclear staining (DAPI, blue), PD-L1 staining (pseudocolored, green), and cytoskeleton staining (phalloidin, red), and the right column (insets) shows PD-L1 expression alone (grayscale) magnified from the region of interest marked by the yellow dotted line. The scale bars are indicated in the micrographs. C Western blots showing the expression of the indicated proteins in IFNγ-stimulated (50 ng/mL, 24 h) HT-29 cells subsequently treated with Mo or Atezolizumab alone or together (Mo + Atezolizumab). The blots are representative of three replicates, and the results are shown in the densitometry graphs. Immunofluorescence images showing the protein levels of D PD-L1 and E active β-catenin in IFNγ-stimulated SW480 cells subsequently treated with Atezolizumab alone or together with Montelukast (Mo + Atezolizumab). Bars, 10 µm. In all western blot panels, GAPDH served as the loading control. MW, relative molecular weight expressed in kilodaltons (kDa). Mean ± SEM. * P < 0.05, *** P < 0.001, two-tailed unpaired t test

Article Snippet: Total β-Catenin (D10A8) XP® Rabbit mAb , BDBioscience , #8480 , 1:5000 , Immunoblotting.

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Control, Molecular Weight, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: The tumor promoter cysteinyl leukotriene receptor 1 regulates PD-L1 expression in colon cancer cells via the Wnt/β-catenin signaling axis

doi: 10.1186/s12964-023-01157-6

Figure Lengend Snippet: Directly antagonizing the Wnt/β-catenin axis negatively impacts PD-L1 expression. A Western blots showing the expression of the indicated proteins in HT-29 CC cells with or without prestimulation (IFNγ 50 ng/mL, 24 h). IFNγ-stimulated cells were further exposed to XAV-939 (Wnt inhibitor, 10 µM) or Atezolizumab alone or both together. The blots are representative of three replicates, and the results are shown in the densitometry graphs. B Immunofluorescence images showing the active β-catenin level in IFNγ-stimulated SW480 cells subsequently treated with XAV-939 (XAV) or Atezolizumab alone or in combination (XAV + Atezolizumab). Bars, 10 μm or 2 µm, as indicated in the images. The magnified images are grayscale images (inset) of active β-catenin in the region of interest marked with the white dotted line. In all the western blot panels, GAPDH served as the loading control. MW, relative molecular weight expressed in kilodaltons (kDa). Mean ± SEM. * P < 0.05, *** P < 0.001, two-tailed unpaired t test

Article Snippet: Total β-Catenin (D10A8) XP® Rabbit mAb , BDBioscience , #8480 , 1:5000 , Immunoblotting.

Techniques: Expressing, Western Blot, Immunofluorescence, Control, Molecular Weight, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: The tumor promoter cysteinyl leukotriene receptor 1 regulates PD-L1 expression in colon cancer cells via the Wnt/β-catenin signaling axis

doi: 10.1186/s12964-023-01157-6

Figure Lengend Snippet: PD-L1 expression positively correlates with CysLT 1 R and β-catenin expression. A Bar graphs showing the relative mRNA expression levels of the indicated transcripts in matched normal and tumor tissues of CC patients ( n = 24). B Western blots showing the expression of the indicated proteins in matched normal and tumor tissues of CC patients ( n = 6). MW, relative molecular weight expressed in kilodaltons (kDa). Pearson correlation plots showing the positive correlation between the transcript levels of CYSLTR1 and CD274 in C TCGA-COAD cohort ( n = 327) and E GSE39582 cohort ( n = 585) of CC patients. Graphs showing CYSLTR1 mRNA expression in CC patients with high or low CD274 mRNA expression in the D TCGA-COAD cohort ( n = 274) and F GSE39582 cohort of CC patients ( n = 585). G Schematic summary showing alterations in the expression of the indicated proteins when CysLT 1 is activated or blocked. The number of patients in each subgroup is shown in the figure. Mean ± SEM. * P < 0.05, *** P < 0.001, Mann‒Whitney U test

Article Snippet: Total β-Catenin (D10A8) XP® Rabbit mAb , BDBioscience , #8480 , 1:5000 , Immunoblotting.

Techniques: Expressing, Western Blot, Molecular Weight