Fig 2: Proliferative breast-associated cells upregulate MCT4 expression upon MCT1 inhibition by AZD3965. (A–D) Cells were assayed in medium containing 25 mmol/L glucose with 10 mmol/L GlutaMAX, 10% FBS and without added lactate. (A) mRNA (left panel) and protein (middle and right panels) expression of MCT1, MCT2, MCT4 and CD147/basigin in T47D cells treated for 24 h ± 10 µM AZD3965 (n = 3–6 for RT-qPCR, n = 3 for WB). (B) As in A, but using MCF7 cells (n = 3–6 for RT-qPCR, n = 6 for WB). (C) As in A, but using MCF10A cells (n = 3–6 for RT-qPCR, n = 3–9 for WB). (D) As in A, but using BJ fibroblasts (n = 3–6 for RT-qPCR, n = 3–6 for WB). All data are shown as means ± SEM. * p < 0.05, ns p > 0.05 compared to vehicle; by Student’s t test (A–D).

Fig 3: Macrophage-specific CD147 deficiency diminishes CD36 expression and may exert other protective effects in atherosclerosis. (A) BMDMs from CD147WT and CD147M–KO treated with or without ox-LDL (50 µg/mL) for 24 h were analyzed by RNA sequencing. The selected genes involved in foam cell formation were graphed in a heatmap. (B) Western blot analysis of LDLR, ABCA1, ABCG1, SR-A, and CD36 in BMDMs isolated from CD147M–KO and CD147M–KI mice and their respective CD147WT littermates incubated with or without ox-LDL (50 µg/mL) for 24 h. (C) Quantification of protein levels of molecules in (B) relative to tubulin. (D) Immunofluorescence staining of CD147, CD36, ABCA1, ABCG1, and LDLR in BMDMs isolated from CD147M–KO and CD147M–KI mice and their respective CD147WT littermates treated with ox-LDL (50 µg/mL) for 24 h. Scale bar, 20 µm. (E) Quantification of the MFI of (D) by means of the Image-Pro Plus 6.0 software. (F,G) GESA GO enrichment analysis for biological process (F) and GESA reactome enrichment analysis (G) are shown as lollipop charts. Statistical significance in gene sets was defined as an FDR < 0.25; NES represents enrichment magnitude and is shown as the size of the point. The -log10 (FDR) of the enrichment is shown on the x-axis; the enriched gene set is shown on the y-axis; and the color of points represent the fraction of gene sets that are significantly upregulated or downregulated. Data represent the mean ± SEM. of n = 3 biologically independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Fig 4: CD147 deficiency in hepatocytes inhibited activation of the NF-?B/NLRP3 signaling pathway. (A) Protein levels of NLRP3 in liver tissues of C57/BL6 NASH model mice (n = 2). Band relative intensity was normalized by a-tubulin, and one-way ANOVA followed by Dunnett’s test was used for statistical analysis. *p < 0.05. (B) mRNA levels of NLRP3 in liver tissues of C57/BL6 NASH model mice (n = 6). One-way ANOVA followed by Dunnett’s test was used for statistical analysis. *p < 0.05 and **p < 0.01. (C) Protein levels of NLRP3 in Bsgflx/flx and Alb;Bsgflx/flx mice fed control or MCD diet (n = 2). (D) mRNA levels of NLRP3 in Bsgflx/flx and Alb;Bsgflx/flx mice fed control or MCD diet (n = 5). One-way ANOVA followed by Bonferroni’s multiple comparison test was used to compare each group. ***p < 0.001. (E) Protein levels of p65 and p-p65 in Bsgflx/flx and Alb;Bsgflx/flx mice fed control or MCD diet (n = 3). One-way ANOVA followed by Bonferroni’s multiple comparison test was used to compare each group. *p < 0.05 and **p < 0.01. (F) Protein levels of nuclear p65 in Bsgflx/flx and Alb;Bsgflx/flx mice fed control or MCD diet (n = 3). One-way ANOVA followed by Bonferroni’s multiple comparison test was used to compare each group. **p < 0.01. (G) mRNA levels of Caspase1 in Bsgflx/flx and Alb;Bsgflx/flx mice fed control or MCD diet (n = 6). One-way ANOVA followed by Bonferroni’s multiple comparison test was used to compare each group. *p < 0.05.

Fig 5: Macrophage-specific CD147 knockin promotes foam cell formation. (A) Generation of macrophage-specific CD147 knockin mice (Lyz2cre/+CD147KIf/f, namely, CD147M–KI mice) is illustrated by the mating scheme. The control WT littermates were Lyz2+/+CD147KIf/f, namely, CD147WT mice. PCR analysis of genomic DNA showed the genotyping. The fragments from top to bottom are the KI Cre gene, WT Cre gene, floxed CD147KI gene and WT CD147KI gene. Genomic DNA from Lyz2cre/+ mice was used as the P.C. for Cre analysis. Genomic DNA from CD147KIf/+ mice were used as the P.C. for CD147 analysis. H2O was used as the N.C. (B) Characterization via western blotting and RT-PCR of BMDMs with or without ox-LDL treatment isolated from CD147WT or CD147M–KI mice. (C) Representative images of Oil Red O staining of CD147WT or CD147M–KI BMDMs that were incubated with or without ox-LDL (50 µg/mL) for 24 h. The scale bar is 50 µm. (D) For quantification, Oil Red O absorbance was measured at 492 nm after extraction with isopropanol. (E) BMDMs from CD147WT or CD147M–KI mice stimulated with or without ox-LDL (50 µg/mL) for 24 h were stained with Bodipy (lipids, green), F4/80 (macrophages, red), and DAPI (nuclei, blue) and examined via confocal microscopy. The scale bar is 20 µm. (F) The total cholesterol and cholesteryl ester contents were determined with a coupled enzyme assay. (G) Representative images of Filipin staining of BMDMs from CD147WT or CD147M–KI mice treated with or without ox-LDL (50 µg/mL) for 24 h. The scale bar is 100 µm. Data represent the mean ± SEM. of n = 3 biologically independent experiments. *P < 0.05, **P < 0.01.