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. 2020 Sep 10;21(18):6613. doi: 10.3390/ijms21186613

Figure 1.

Figure 1

C-P4H1 is expressed and purified from HEK-293FT cells. (A) Expression of C-P4H1 was analyzed by western blot with anti-P4H1α and anti-flag antibody. Cell lysates were collected from control and 293 FT cells transfected with P4HA1 and P4HB constructs. (B) Ponceau staining showed the expression and purification of C-P4H1 from 293 FT cells and CHO cells. The letters indicate: L (Total cell lysates with HGLB), Un (unbinding samples after cell lysates incubated with M2 gel), W (The last time washed sample in NET2), E1 (The first time elution sample in 0.25 µg/µL 3× Flag), E2 (The second time elution sample in 0.25 µg/µL 3× Flag), Gel (The remaining sample on M2 gel after elution). (C) Western blot analysis of C-P4H1 expression and purification with anti-P4Hα. (D) Purified P4H1 samples was analyzed by 8% Native PAGE gel with Coomassie Blue staining; BSA was included as a control.