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A, B
Cell proliferation (n = 3, A) and viability (n = 3, B) assays on the effect of Orlistat (100 μM) on cells at indicated doses for 72 h (upper and lower panels respectively). Significance in differences in proliferation and viability indexes, in which vehicle acted as control, was determined by t‐test. Error bars denote SEM.
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C
Western blot analysis of EGFR, FASN, Bax, Bak, and survivin protein expression from cells exposed to either Gefitinib or Orlistat for 48 h. ACTB is selected as a housekeeping gene. Vehicle‐treated cells acted as controls.
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D
Western blot data showing EGFR palmitoylation in Orlistat‐treated cells (48 h). Hydroxylamine (HAM), a strong reducing agent that cleaves palmitate from cysteine residues, is necessary for biotinylation. The omission of HAM cleavage (HAM‐) serves as negative control for the ABE assay.
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E
Western blot images showing ubiquitination status of EGFR in cells after exposure to vehicle, Gefitinib or Orlistat for 48 h.
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F
FACS analysis (n = 2) for Annexin V/PI staining shows percentages of early (Annexin+) and late (Annexin and PI+) apoptotic cells treated with vehicle, Gefitinib or Orlistat for 48 h.