Fig. 7: JAB1 and CUL4B enhance the proliferation and invasion of breast cancer cells, and induce fatty acid metabolic reprogramming, through the inhibition of PPARG/ACSL5 expression.

A The protein expression levels of ACSL5 and PPARG were detected in MDA-MB-231 cells with JAB1 knockdown or overexpression (left panel). The protein expression levels of ACSL5 and PPARG were also detected in MDA-MB-231 cells with CUL4B knockdown or overexpression (right panel). B The efficiency of ACSL5 or PPARG knockdown in MDA-MB-231 cells was confirmed using qRT-PCR. C Clone formation assays were conducted in MDA-MB-231 cells after transfection with control, shJAB1, shCUL4B, siPPARG, or siACSL5. D Transwell assays were conducted on MDA-MB-231 cells with control, shJAB1, shCUL4B, siPPARG, or siACSL5 transfection. E The relative levels of serum-free free fatty acids (FFA) and serum-free triglycerides (TG) were detected in MDA-MB-231 cells transfected with control, shJAB1, shCUL4B, siPPARG, or siACSL5. F, G Wide-targeted lipidomics analysis revealed upregulated lipid species and their distribution in MDA-MB-231 cells with JAB1 overexpression, CUL4B overexpression, and ACSL5 knockdown. H Wide-targeted lipidomics analysis results revealed the levels of arachidonic acid (AA) in each group. I Clone formation assays were performed in MDA-MB-231 cells (shSCR, shJAB1, and shCUL4B groups) following supplementation with 15 μM arachidonic acid. J Transwell assays were conducted in MDA-MB-231 cells (shSCR, shJAB1, and shCUL4B groups) following supplementation with 15 μM arachidonic acid. K Sphere formation assays were performed in MDA-MB-231 cells (shSCR, shJAB1, and shCUL4B groups) following supplementation with 15 μM arachidonic acid. L The expression levels of metabolites involved in fatty acid oxidation were detected by qRT-PCR in MDA-MB-231 cells. M Mitochondrial oxygen consumption rate (OCR) was examined in MDA-MB-231 cells transfected with control, shJAB1, shCUL4B, siPPARG, or siACSL5. J1, JAB1; C4B, CUL4B; PG, PPARG; A5, ACSL5. The error bars indicate the mean ± SD derived from three separate experiments; two-tailed unpaired t-test, *p < 0.05, **p < 0.01, ***p < 0.001.