Journal: eBioMedicine

Article Title: Macrophage CD5L is a target for cancer immunotherapy

doi: 10.1016/j.ebiom.2023.104555

Figure Lengend Snippet: Lung and liver cancer cell-conditioned media (CM) induce an IL10-like phenotype and CD5L expression in macrophages. a) Principal Component Analysis (PCA) scatterplot of PB monocytes treated for 72 h with medium alone (control), reference activation stimuli (IFN/LPS, IL4, and IL10), or cancer cell-CM. Projection based on the expression profile of surface markers characterized by multicolor flow cytometry. PC1 and PC2 represented in each axis depict the first and second principal components, respectively. b) Expression of CD80 , CD163 , CD206 , VEGF , MERTK and c) CD5L mRNA was assessed by RT-qPCR in PB monocytes treated for 24 h with the indicated stimuli. mRNA levels normalized to GAPDH, and fold induction levels were calculated using the average expression of each gene in control macrophages as a reference. Data are represented as mean ± SEM (n = 5 to 9). d) CD5L immunofluorescence staining (green) in macrophages treated with the indicated stimuli for 72 h. Nuclei were counterstained with Hoechst 33258 (blue). Scale bar represents 20 μm. CD5L mean fluorescence intensity (MFI) was calculated with ZenLite software and is represented as MFI ± SEM of 50 macrophages scored in random fields (right) (n = 3). Significance was calculated using the Mann–Whitney t-test (∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001).

Article Snippet: CD5L fluorescence intensity was quantified using ZenLite software (Carl Zeiss Microscopy GmbH).

Techniques: Expressing, Activation Assay, Flow Cytometry, Quantitative RT-PCR, Immunofluorescence, Staining, Fluorescence, Software, MANN-WHITNEY