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. 2022 Dec 16;11(24):4101. doi: 10.3390/cells11244101

Figure 4.

Figure 4

KLK6 is responsible for macrophage-promoted CXCL1 production in cancer cells. (A) RAW 264.7 cells were transfected with the mock vector (Mock) or GFP-tagged KLK6 expression vector (pCMV3-KLK6). KLK6 protein expression in CM of mock- or pCMV3-KLK6-transfected cells was analyzed by Western blot. (B,C) B16F10 cells were treated with the CMs of mock- or pCMV3-KLK6-transfected cells for 24 h. (B) The mRNA levels of the CXCL1 and CXCL2 in the indicated CM-treated B16F10 cells were analyzed by RT-PCR. (C) The protein levels of CXCL1 and CXCL2 in the supernatants of the indicated CM-treated B16F10 cells were analyzed by ELISA. (D) RAW 264.7 cells were transfected with the control or two different KLK6 siRNAs. KLK6 mRNA levels were analyzed by RT-PCR. The KLK6 secreted in CMs was analyzed by Western blot (right panel). (E,F) B16F10 cells were treated with the CMs of RAW 264.7 cells transfected with the indicated siRNA for 24 h. (E) The mRNA levels of CXCL1 and CXCL2 in the indicated CM-treated B16F10 cells were analyzed RT-PCR. (F) The protein levels of CXCL1 and CXCL2 in the supernatants of the indicated CM-treated B16F10 cells were analyzed by ELISA. Statistical significance was determined by the Student’s t-test. * p < 0.05. Data are representative of three experiments.