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. 2021 Aug 6;22(16):8481. doi: 10.3390/ijms22168481

Figure 4.

Figure 4

BCL2L15/Bfk is selectively expressed in HT-29 and SW620 CRC cells and it is downregulated upon YY1 silencing. (A) q-RT-PCR analysis of BCL2L15 expression in the four CRC cell lines (GAPDH used as housekeeping); data are expressed as 2−ddCt and compared with SW620 normalized expression. (B) Immunoblot of Bfk (17 KDa) and β-Actin (42 KDa) proteins expression in the four CRC cell lines. (C) Densitometry analysis of Bfk expression (normalized to β-Actin). (D) q-RT-PCR analysis of YY1 expression in HT-29 (CTRL, KD-01, KD-02), GAPDH used as housekeeping, data are expressed as 2−ddCt compared to CTRL. (E) q-RT-PCR analysis of BCL2L15 expression in HT-29 (CTRL, KD-01, KD-02), GAPDH used as housekeeping, data are expressed as 2−ddCt compared to CTRL. (F) Immunoblot and densitometry of HT-29 (CTRL, KD-01, KD-02). Signal detected for YY1, Bfk and β-Actin. (G) q-RT-PCR analysis of YY1 expression in SW620 (CTRL, KD-01, KD-02), GAPDH used as housekeeping, data are expressed as 2−ddCt compared to CTRL. (H) q-RT-PCR analysis of BCL2L15 expression in SW620 (CTRL, KD-01, KD-02), GAPDH used as housekeeping, data are expressed as 2−ddCt compared to CTRL. (I) Immunoblot and densitometry of SW620 (CTRL, KD-01, KD-02). Signal detected for YY1, Bfk and β-Actin. (J) Immunoblot and densitometry of HT-29 and SW620 (CTRL, KD-01, KD-02) treated with 30µM 5-FU, from 0 to 72 h. Signal detected for YY1, Bfk, c-Casp-3 and β-Actin. Values are presented as Mean  ±  SD. ** p < 0.01; *** p < 0.001; **** p < 0.0001.