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. 2021 Sep 10;13(18):4552. doi: 10.3390/cancers13184552

Figure 2.

Figure 2

MT2A promotes the expression and secretion of IGFBP2 in CAF-like cells. (A,B) Three types of CAF-like cells (CAF8. CAF9, and CAF15) were transfected with siRNA targeting MT2A (siMT2A) and control siRNA (siNC). The knockdown was confirmed by qRT-PCR (A) and Western blotting (B). After Western blotting, the normalized relative expression fold-change was calculated using the ImageJ software, and the values were arbitrarily set as 1.00 for cells transfected with siNC. (C) Antibody array with monocultured MSC (monoMSC), CAF15, siNC-transfected CAF15 and siMT2A-transfected CAF15 conditioned medium. The intensity of insulin-like growth factor binding protein 2 (IGFBP2) was quantified using the public domain software ImageJ, normalized to a positive control reference spot. (D–F) Expression levels and secretory concentrations of IGFBP2 in CAF8, CAF9, and CAF15 cells were compared with those in MSCs using qRT-PCR (D), ELISA (E), and Western blotting (F). After Western blotting, the normalized relative expression fold-change was calculated using the ImageJ software, and the values were arbitrarily set as 1.00 for control MSCs. (G) Expression of IGFBP2 was detected by qRT-PCR in CAF8, CAF9, and CAF15 cells transfected with siNC and siMT2A. (H) The secretory concentrations of IGFBP2 were detected by ELISA in CAF8, CAF9, and CAF15 cells transfected with siNC and siMT2A. (I) Western blotting for IGFBP2 in CAF8, CAF9, and CAF15 cells transfected with siNC and siMT2A. After Western blotting, the normalized relative expression fold-change was calculated using the ImageJ software, and the values were arbitrarily set as 1.00 for cells transfected with siNC. Data are presented as mean ± SEM (* p < 0.05, ** p < 0.01, *** p < 0.001).