Skip to main content
. 2023 Oct 23;14:6690. doi: 10.1038/s41467-023-42332-0

Fig. 3. TAMs promote the stabilization of FGL1 by activating NF-κB in the liver microenvironment.

Fig. 3

A UMAP embedding of transcriptional profiles from a previously published single-cell CRLM atlas (No. of cells = 98,072). B Stacked bar chart showing the proportion of cells in each cluster in primary normal tissue (PN), primary tumor tissue (PT), and liver metastatic tissue (LM). C UMAP embedding and relative abundance of myeloid cells in PNs (n = 10), PTs (n = 10), and LMs (n = 10). D IB detection of FGL1 expression in indicated cells cocultured with TAMs for 16 h. TAMs, tumor-associated macrophages. E qRT‒PCR analysis of FGL1 mRNA levels in indicated cells incubated with TAM, n = 3 biologically independent experiments. F qRT‒PCR analysis of FGL1 mRNA levels in MC38 cells sorted from primary and liver metastatic tumor tissues, n = 5 biologically independent experiments. G IB detection and quantification of FGL1 expression in MC38 cells stimulated with the indicated cytokines (10 ng/mL), n = 3 biologically independent experiments. H IB detection and quantification of FGL1 expression in indicated cells incubated with TAM conditioned medium (TAM-CM) following treatment with 40 μg/mL cycloheximide (CHX) for the indicated times, n = 3 biologically independent experiments. I IB detection of FGL1 expression in MC38 cells incubated with TAM-CM and treated with the indicated inhibitors (10 μM) for 1 h. J IB detection of FGL1 expression in indicated cells incubated with TAM-CM. K IB detection of Flag-FGL1 expression in WT or p65−/− MEFs transiently overexpressing Flag-FGL1 following incubation with TAM-CM. L Coimmunoprecipitation analysis of the interaction between HA-Ub and Flag-FGL1 in indicated cells pretreated with Baf-A1 (12 h, 100 nM) and subjected to TAM-CM. M IB detection and quantification of Flag-FGL1 expression in indicated cells, n = 3 biologically independent experiments. The data in (C) are presented as a box-and-whisker graph (min-max), and the horizontal line across the box indicates the median. IB experiments in (D, G–M) were repeated three times, the data are representative of three biologically independent experiments. The data in (E–G, H, M) are presented as the mean ± SD. P values were determined by one-way ANOVA (C, E, G) and two-tailed unpaired Student’s t test (F, H, M).