Fig. 6: Inhibition of PP1 phosphatase activity promotes anchorage-independent lung cancer cell growth. | npj Precision Oncology

Fig. 6: Inhibition of PP1 phosphatase activity promotes anchorage-independent lung cancer cell growth.

From: IKKα promotes lung adenocarcinoma growth through ERK signaling activation via DARPP-32-mediated inhibition of PP1 activity

Fig. 6

a Human lung cancer HCC827 cells treated with calyculin A (50 nM) for the indicated times were lysed using 1× RIPA buffer supplemented with protease and phosphatase inhibitors. An equal amount of protein was separated with 4–20% SDS-PAGE and transferred to polyvinyl difluoride membranes. Antibody-reactive protein bands were visualized after overnight incubation of primary antibodies against phosphorylated PP1α (Thr320), total PP1α, PARP-I, phosphorylated ERK (Thr202/Tyr204), total ERK, and α-tubulin (loading control). b Human NSCLC HCC827, PC9, and H1650 cells were incubated with vehicle (DMSO) or calyculin A (50 nM) for 15 min prior to plating. Representative images indicate cell colonies grown on the soft-agar cell culture dishes after 2 weeks of incubation. Scale bar 200 µm. c Bar graphs show the average count of cell colonies observed after 2 weeks of incubation. Bar graphs indicate mean ± SEM (n = 9). A value of P ≤ 0.05 was considered significant, two-tailed unpaired t-test.

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