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. 2020 Mar 10;39(18):3754–3773. doi: 10.1038/s41388-020-1251-2

Fig. 4. GPRC5A deficiency promotes the phosphorylation and acetylation of NF-kB and mediates the elevation of ABCG1 expression in AT2 cells.

Fig. 4

a Top ten upregulated pathways in KO:AT2 vs WT:AT2 cells by KEGG pathway enrichment analysis; b Heatmap clustering of the global pattern of ATP binding cassette (ABC) gene expression in KO:AT2 and WT:AT2 cells conducted using the hierarchical clustering (HCL) algorithm; qPCR (c) and WB (d) analysis of ABCG1 in isolated AT2 cells from KO or WT mice; e IF analysis of ABCG1 in the S/TB region of lung tissue from WT or KO mice, (Bar = 100 µm); Illustration (f), WB analysis of knockdown efficiency (g), colony formation (h) and tumor initiation (i) of KO:AT2 cells with abcg1 knockdown cocultured with KO:Lgr5 cell; j WB analysis of phosphorylated-p65 (p-p65), total p65 (t-p65) in AT2 cells from WT or KO mice; k qPCR analysis of the mRNA level of ABCG1 in KO-AT2 cells treated with PS1145 (inhibitor of NF-kB), C646 (inhibitor of p300) or PS1145 combined with C646 and then cocultured with KO:Lgr5; l WB analysis of the phosphorylation and acetylation of NF-kB p65 and ABCG1 in KO:AT2 cells treated with PS1145, JSH-23, C646 or AA and then cocultured with KO:Lgr5; m WB analysis of the phosphorylation and acetylation of NF-kB p65 and ABCG1 in α6 knockdown or β4 knockdown or combined knockdown in KO:AT2 cells cocultured with KO:Lgr5 cells; n WB analysis of the phosphorylation and acetylation of NF-kB p65 and ABCG1 in KO:AT2 cells that cocultured with ECM1-WT or ECM1-MT KO:Lgr5; o Co-IP analysis of p65 and p300 in KO:AT2 cells that cocultured with ECM1-WT or ECM1-MT KO:Lgr5; Data were collected from three independent experiments with triplicate samples. *P < 0.05; **P < 0.01; ***P < 0.001.