Figure 1.

PIGN expression is cell cycle regulated. (A) Schematic of cell cycle synchronization protocols. Cell synchronizations were achieved using serum starvation for 72 h (Go/G1), double-thymidine block (G1/S), or double thymidine block and release followed by nocodazole treatment (G2/M). Cells were collected post-treatment for western blotting, RT-qPCR, and flow cytometry analyses. PIGN was expressed in a cell cycle-dependent manner in multiple cell lines: (B) HL60, (C) K562, (D) KCL22, and (E) JURKAT with suppressed expression from early S-phase to the G2/M phase. All western blot images were cut within the molecular weight ranges of the protein targets prior to hybridization. Images generated from these cut blots by scanning the developed films. Images were cropped and labeled using Adobe Photoshop CC 2017 (version 18). Where applicable, representative full images of the blots are presented in Figure 3 of the supplementary immunoblotting data.