Fig 2: Lrh1 mRNA expression and localization in porcine oocytes and early embryos at various developmental stages. (A) mRNA was collected during different phases of porcine embryo development. (B) Laser scanning confocal microscopy images of liver receptor homolog 1 immunostaining during the MII, 2C, 4C, and blastocyst stages. GV, MII, 1C, 2C, 4C, and BL indicate germinal vesicle, meiosis II, one-cell stage, two-cell stage, four-cell stage, and the blastocyst, respectively. ** indicates statistically significant difference (P < 0.01) from GV. Data are presented as the mean (± standard deviation of the mean) of three independent experiments.

Fig 3: PGC1α is required for maintenance of lung identity in BRAFV600E driven tumors.(A) Tumors were induced in cohorts of BrafCAT/+ and BrafCAT/+;Ppargc1af/f mice via intranasal instillation of 106 PFU Ad5-SpC-CRE and harvested from each genotype 12 weeks post tumor induction via tissue dissociation and FACS. GSEA analyses of hallmark pathways and lung identity gene sets. Black bars indicate adjP <.05, gray bars indicate Benjamini-Hochberg corrected enrichment statistic adjP ≥. 05. (B) Immunostaining confirms decreased expression of the AT2 markers SFTPA and LYZ in BRAFV600E/PGC1αNULL tumors. (C) Quantitation demonstrating a significant decrease of LYZ immunoreactivity in BRAFV600E/PGC1αNULL tumors. Wilcoxon rank sum p val. = 0.0288. (D) Luciferase assays in HEK293T cells demonstrating the cooperation of NKX2-1, FOXA1, PGC1α, and NR5A2 in transactivation of surfactant promoters. All three promoters showed significant induction by ordinary one-way ANOVA (p<0.0001). Comparison of individual groups to mock transfected controls by Dunnett’s test for multiple comparisons: (*) p=0.0189, (****) p<0.0001. (E) Co-Immunoprecipitation of NKX2-1 by immunoprecipitation with a mouse monoclonal antibody recognizing PGC1α but not with IgG. (F) Co-Immunoprecipitation of PGC1α by immunoprecipitation with a mouse monoclonal antibody recognizing NKX2-1 but not with mouse IgG.10.7554/eLife.43668.038Figure 7—source code 1.R script to perform gene set enrichment analysis on Figure 7—source data 1, as well as plot these results.10.7554/eLife.43668.039Figure 7—source code 2.R script to perform statistics on Figure 7—source data 2, as well as plot these results eLife’s transparent reporting form.10.7554/eLife.43668.040Figure 7—source data 1.DEseq2 output of differentially expressed genes comparing BRAFV600E/PGC1αNULL and BRAFV600E/PGC1αHET driven tumors.10.7554/eLife.43668.041Figure 7—source data 2.Cellprofiler output quantifying immunofluorescence of LYZ in BRAFV600E/PGC1αNULL and BRAFV600E/PGC1αWT driven tumors.10.7554/eLife.43668.042Figure 7—source data 3.Data from luciferase assays looking for transactivation of Sftpa, Sftpb, and Sftpc promoters.

Fig 4: The effect of liver receptor homolog 1 (LRH1) inhibition on the embryo development. The parthenotes were treated with the specific LRH1 antagonist 505601 in concentrations of 50 or 100 μM. (A) The percentages of those in the two-cell, four-cell, blastocyst, and hatching blastocyst stages are shown by bars. (B) The images of embryos treated with different concentrations of 505601 are shown. (C) Expression of Fn1, Itgα5, and Cox2 mRNA in hatching blastocysts in the presence or absence of the LRH1 antagonist. Statistical significance of the differences from the control group is indicated as follows: * P < 0.05, ** P < 0.01, and *** P < 0.001. Data are presented as the mean (± standard deviation of the mean) of three independent experiments.

Fig 5: LRH1 expression in non‐small cell lung cancer (NSCLC) tissue specimens. Representative (a) quantitative real‐time PCR and (b) Western blot analysis of LRH1 expression in 16 paired NSCLC clinical tissue specimens. Histograms of LRH1 messenger RNA (mRNA) expression and pooled data in adjacent normal lung (N, n = 16) and NSCLC (T, n = 16) tissues. The relative expression of LRH1 mRNA and LRH1 were highly expressed in NSCLC tumor tissues compared to normal lung tissues. Data are expressed as mean ± standard deviation (SD). *P < 0.05. (c) Representative immunohistochemistry staining images of samples from an NSCLC patient (T) and adjacent normal lung (N) tissues showed LRH1 expression in NSCLC tissues. The expression of LRH1 protein significantly increased in tumor tissues compared to adjacent normal lung tissues. A histogram of pooled data (right) of NSCLC (T, n = 156) and normal lung (N, n = 30) tissues. The percentage of NSCLC tissues with high LRH1 expression significantly increased compared to that of normal lung tissues. *P < 0.05.