Fig. 6: OFD1 sustains tumor cell proliferation and tumor growth.
a Proliferation (Upper Panel) and immunoblot (Lower Panel) analyses of indicated cell lines expressing Doxycycline-inducible control shRNA or OFD1 shRNA. Data shown represented as mean values mean ± SD, error bar was defined as SD. b Proliferation assay of indicated cell lines expressing control shRNA or OFD1 shRNA. Cells were stained with crystal violet. c Quantitation of mitotic events in the indicated cell lines with or without knockdown of OFD1. 300 cells examined over three independent experiments, data shown represented as mean values ± SD, error bar was defined as SD, two-tailed unpaired student’s t-test. d Quantification of the percentages of cell viability upon OFD1 peptide treatment as indicated for 7 days. 300 cells examined over three independent experiments, P = 0.0126; P = 0.0015; P = 0.0003; P = 0.0002; P = 0.0012; P = 0.0001, two-tailed unpaired student’s t-test. Data shown represented as mean values mean ± SD, error bar was defined as SD. e Proliferation assay of indicated cell lines expressing OFD1 shRNA. Left Panel: Cells were grown in the absence of Doxycycline for 10 days. Cells were grown in the presence of Doxycycline for 6 days, DOX was then removed and the cells grown for 20 days. Cells were visualized by crystal violet staining. f Tumor volume of xenografts formed after subcutaneous injection of NOD/SCID mice with PANC-1 cells expressing control shRNA or OFD1 shRNA in the presence or absence of Doxycycline in the animal diet. Tumor size was measured twice per week. 9 mice per group per time point examined over three independent experiments, two-tailed unpaired student’s t-test. Data shown represented as mean values ± SD, error bar was defined as SD. g Weight of PANC-1 xenograft tumors from experiments shown in f. Data shown represented as mean values ± SD, error bar was defined as SD, P = 3 × 10−6; P = 3 × 10−6, two-tailed unpaired student’s t-test. h Tumor images of indicated PANC-1 xenograft tumor genotype from experiments shown in (f). i Tumor volume of xenografts formed after subcutaneous injection of NOD/SCID mice with HT-29 cells infected with an indicated Doxycycline-inducible lentivirus encoding control shRNA or OFD1 shRNA in the presence or absence of Doxycycline in the animal diet. The tumor sizes were measured twice per week. 12 mice per group per time point examined over three independent experiments, two-tailed unpaired student’s t-test. Data shown represented as mean values ± SD, error bar was defined as SD. j Weights of HT-29 xenograft tumors from experiments in (i). Data shown represented as mean values ± SD, error bar was defined as SD, P = 1.3 × 10−9, two-tailed unpaired student’s t-test. k Tumor images of indicated HT-29 xenograft tumor genotype from experiments shown in i. l Tumor volume of xenografts formed after subcutaneous injection of NOD/SCID mice with MDA-MB-231 cells expressing Dox-inducible OFD1 shRNA in the presence or absence of Doxycycline in the animal diet. The tumor sizes were measured twice per week. Eight mice per group per time point. 8 mice per group per time point examined over three independent experiments, two-tailed unpaired student’s t-test. Data shown represented as mean values ± SD, error bar was defined as SD. m Weights of MDA-MB-231 xenograft tumors from experiments in (l). Data shown represented as mean values ± SD, error bar was defined as SD, P = 4 × 10−6, two-tailed unpaired student’s t-test. n Tumor images of indicated MDA-MB-231 xenograft tumor genotype from experiments shown in (l).
