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. Author manuscript; available in PMC: 2019 Dec 5.

Published in final edited form as: Mol Cell Endocrinol. 2018 Jun 2;477:57–69. doi: 10.1016/j.mce.2018.06.001

Fig. 2 — Transient co-expression of LIP and c-Jun synergize to induce expression from the PR promoter-reporter. Transient co-transfection of C/EBPβ and c-Jun expression vectors were carried out in duplicate and repeated at least three times in murine mammary tumor cell lines of epithelial origin with the −2494/+769 PR promoter-reporter. (A, B, C, D) MC7-L1 cells. The varied ng quantities of the expression vector per 5×10⁴ cells are indicated below the x-axis. For panels A, B, and C, C/EBPβ expression vectors were co-transfected with 100 ng of c-Jun expression vector in some transfections. Luminometer values were normalized for expression from a co-transfected SV40 early enhancer/promoter-Renilla luciferase reporter. These values were then normalized to a relative value of 1.0 for cells receiving only “empty” pcDNA3.1 and neither C/EBPβ nor c-Jun expression vector. The data presented are the means of at least three experiments with SEM. p was calculated by Student’s t-Test in comparison to the control transfection for transfections involving a single transfected expression vector, and in comparison to transfections involving a C/EBPβ vector alone for co-transfections involving both C/EBPβ and c-Jun expression vectors. *, p < 0.1; **, p < 0.05; ***, p < 0.01. (E) MC4-L2 cells. 100 ng LIP and 100 ng c-Jun expression vectors per 5×10⁴ cells. Values were normalized and statistics performed as described above. (F) MC4-L3 cells. 50 ng LIP and 50 ng c-Jun expression vectors per 5×10⁴ cells. Values were normalized and statistics performed as described above. (G) A western blot confirms similar levels of C/EBPβ expression among the various isoforms in transfection of MC7-L1 cells. c-Jun is overexpressed in cells transfected for c-Jun expression. β-tubulin was detected as a loading control.

Fig. 2 — Transient co-expression of LIP and c-Jun synergize to induce expression from the PR promoter-reporter. Transient co-transfection of C/EBPβ and c-Jun expression vectors were carried out in duplicate and repeated at least three times in murine mammary tumor cell lines of epithelial origin with the −2494/+769 PR promoter-reporter. (A, B, C, D) MC7-L1 cells. The varied ng quantities of the expression vector per 5×10⁴ cells are indicated below the x-axis. For panels A, B, and C, C/EBPβ expression vectors were co-transfected with 100 ng of c-Jun expression vector in some transfections. Luminometer values were normalized for expression from a co-transfected SV40 early enhancer/promoter-Renilla luciferase reporter. These values were then normalized to a relative value of 1.0 for cells receiving only “empty” pcDNA3.1 and neither C/EBPβ nor c-Jun expression vector. The data presented are the means of at least three experiments with SEM. p was calculated by Student’s t-Test in comparison to the control transfection for transfections involving a single transfected expression vector, and in comparison to transfections involving a C/EBPβ vector alone for co-transfections involving both C/EBPβ and c-Jun expression vectors. *, p < 0.1; **, p < 0.05; ***, p < 0.01. (E) MC4-L2 cells. 100 ng LIP and 100 ng c-Jun expression vectors per 5×10⁴ cells. Values were normalized and statistics performed as described above. (F) MC4-L3 cells. 50 ng LIP and 50 ng c-Jun expression vectors per 5×10⁴ cells. Values were normalized and statistics performed as described above. (G) A western blot confirms similar levels of C/EBPβ expression among the various isoforms in transfection of MC7-L1 cells. c-Jun is overexpressed in cells transfected for c-Jun expression. β-tubulin was detected as a loading control.