Fig. 6: Regulation of eDHFR-Luciferase expression in vivo using IP OVCAR8 cell tumor model.
From: Regulation of eDHFR-tagged proteins with trimethoprim PROTACs
A Schematic of eDHFR-Luciferase direct fusion construct. Luciferase was directly fused to the C-terminus of eDHFR to allow for regulation of the luminescent protein with 7c. B OVCAR8-eDHFR-Luciferase cells were incubated with 7c for 4, 8, 12, 24, and 48 h. Representative data from n = 3. C 10 × 106 OVCAR8-eDHFR-Luc cells were injected intraperitoneally (IP) in CD-1 nu/nu mice. Following 4 weeks of tumor growth, BLI was performed to measure baseline Luciferase expression (Day −4). On Day 0, 200 mg/kg of 7c (or vehicle control) was administered IP 3 times every 3 h, and BLI was performed 1 h after the last dose and monitored for 9 days. n = 5 for each group, data points are mean ± SEM. Animals from the D-4, D0, and D3 are shown. D Quantification over time of average BLI signal for mice given 200 mg/kg of 7c compared to vehicle control as described in (C). E Percent (%) of maximum luminescence from BLI on Day 3 post-treatment. Data was normalized to baseline luminescence signal on Day 0 and groups were compared using a two-tailed unpaired t test. **p-value = 0.0093. n = 5 for each group, data points are mean ± SEM.
