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. 2020 Feb 11;10:2309. doi: 10.1038/s41598-020-59028-w

Figure 1.

Figure 1

Doxycycline regulated expression of recombinant DDR1b and DDR2 in HT1080 cells. HT-DDR1b (A) and HT-DDR2 (B) cells processed to express or repress recombinant DDRs, as described in the Methods section, were incubated in serum-free media supplemented with or without 20 μg/ml of COL1 for various times at 37 °C. At the end of the incubation period, the cells were lysed in RIPA buffer and equal protein amounts per lane (30 µg) were resolved by reducing SDS-PAGE followed by immunoblot analyses using antibodies to phosphorylated DDR1b (Y513) (A, middle panel) or DDR2 (Y740) (B, middle panel). The same blots were then stripped and reprobed with antibodies against total DDRs (upper panels), and against β-actin, as loading control (lower panels). Control (Ctrl.): cells (harvested at 48 h), which were incubated with or without DOX but not stimulated with COL1. CTF, C-terminal fragment of DDR1b; pCTF, pDDR1b, and pDDR2 refers to the phosphorylated protein. Full-length blots are presented in Supplementary Fig. 9.