SCID (severe combined immunodeficient) beige mice (CB17.Cg-PrkdcscidLystbg-J/Crl), aged 4 to 6 weeks, purchased from Charles River, were injected subcutaneously with 5 × 106 cells in 1:1 solution with Matrigel in both flanks. Mice with established A549-GFP-Luciferase tumors (100 mm3) were treated intraperitoneally each day with doxycycline (20 μg in 0.5-cm3 water) to induce expression of shRNA targeting either a control scramble sequence or DHHC20. Tumors were measured on days −2, 0 (day of doxycycline addition), 5, and 8 using in vivo bioluminescent imaging. To image, mice were anesthetized and intraperitoneally injected with d-luciferin (GoldBio, LUCNA-1G) in PBS at 150 mg/kg. Luminescent signals were acquired 15 min after injection with the IVIS Spectrum (Caliper Life Sciences). Analysis was performed using Living Image 4.5 (PerkinElmer).

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