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. 2021 Mar 13;40(15):2725–2740. doi: 10.1038/s41388-021-01740-6

Fig. 7. Lactate activates CAFs.

Fig. 7

A Conditioned medium (CM) from 24 h culture of NAF823, MCF7, and MCF7-POU1F1 cells was added to NAF823 for 24 h and α-SMA and β-actin protein levels were evaluated by qWB. B qWB of POU1F1 and β-actin in MCF7 and MCF7-POU1F1 cells. C qWB of α-SMA and β-actin in NAF823 after administration of CM-NAF823, CM-MCF7 and CM-MCF7-POU1F1 for 24 h, and representative histograms from two independent qWB. D CM from 24 h culture of NAF823, control MDA-MB-231 (MDAsgC), and MDA-MB-231 cells after POU1F1 knockout (MDAsgPOU1F1) was added to NAF823 for 24 h and α-SMA and β-actin were evaluated by qWB. E qWB of POU1F1 and β-actin in MDAsgC and MDAsgPOU1F1 cells. F qWB of α-SMA and β-actin in NAF823 after CM-NAF823, CM-MDAsgC, and CM-MDAsgPOU1F1 treatments, and representative histograms from two independent qWB. G–H Western blot of α-SMA and β-actin in NAF823 and normal mammary fibroblasts (NMFs) treated with vehicle (0) and lactate (Lac, 20 mM) for 24 h. Data are expressed as mean ± SEM from three independent quantitative western blots. **P < 0.01, and ***P < 0.001. I LDHA and ACTA2 mRNA expression significantly correlate with relapse-free survival (RFS) in human breast tumor samples (http://kmplot.com). J Model of POU1F1-induced metabolic reprogramming and cancer progression. POU1F1 transcriptionally regulates LDHA leading pyruvate into lactate, which in turn increases proliferation, migration, and invasion of breast cancer cells and induces fibroblast activation.