Fig. 5: MARK3 is activated by metabolic stress inducers. | Communications Biology

Fig. 5: MARK3 is activated by metabolic stress inducers.

From: The metabolic stress-activated checkpoint LKB1-MARK3 axis acts as a tumor suppressor in high-grade serous ovarian carcinoma

Fig. 5

a Immunoblotting of chemical screening for metabolic stress inducers. Among tested compounds, anisomycin and thapsigargin increase the kinase-activated phosphorylation of doxycycline (DOX)-inducible MARK3. b Immunoblotting of chemical screening for DNA damage inducers with anisomycin as a positive control. DNA damage does not directly increase the phosphorylation of DOX-inducible MARK3. c Immunoblotting of inhibition experiments for the upstream regulators of MARK3. LKB1 knockdown interferes with DOX-inducible MARK3 activation in response to anisomycin treatment. d CRISPR–Cas9 experiment for DOX-inducible MARK3 knockout in TYK-nu cells. TYK-nu is a high-grade serous ovarian carcinoma (HGSOC)-like cell line, harboring the oncogenic TP53 R175H mutation. The copy numbers of LKB1, TAOK1, PIM1, and MARK3 are duplicates, suggesting that MARK3 can become active in TYK-nu. The genomic profile of TYK-nu is sourced from the Cancer Cell Line Encyclopedia (CCLE). e, f Cell viability assay for DOX-inducible MARK3 knockout in TYK-nu. The cytotoxic effects of anisomycin (e) and thapsigargin (f) are amplified upon MARK3 knockout. Relative cell number shows the relative ratio between the number of cells in the absence of the inhibitors (0 nM = 1.0). and the number of cells in the presence of various concentrations of anisomycin or thapsigargin. Error bars represent mean ± standard deviation (SD) of three biological replicates. Statistical analysis was performed using unpaired Student’s t-test (Anisomycin 250 nM; Thapsigargin 100 nM; *P < 0.01).

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