Journal: Cell reports

Article Title: SMAD4 represses FOSL1 expression and pancreatic cancer metastatic colonization

doi: 10.1016/j.celrep.2021.109443

Figure Lengend Snippet: (A) Immunoblot of SMAD4 gene deletion in SMAD4-WT PDAC cell lines (PANC1 and PATU8902) and SMAD4 re-expression in SMAD4 null PDAC cell lines (HPAC and PANC0327). (B) qRT-PCR of SERPINE1 mRNA expression in SMAD4 isogenic PDAC cell lines treated for 24 h with 10 ng/mL TGF-β. Mean ± SD of three biological replicates is shown. (C) Longitudinal quantification of tumor volume in mice orthotopically implanted with LacZ- or SMAD4-expressing HPAC cells with bioluminescence imaging (BLI) (red indicates HPAC SMAD4; blue indicates HPAC LacZ). Total flux was measured weekly during 42 days (n = 8 for each genotype; standard deviation is reported in error bars). (D) Ultrasound reported tumor volume in mice orthotopically implanted with LacZ- or SMAD4-expressing HPAC cells at 42 days after injection (n = 8 for each cohort; standard deviation is reported in error bars) (E) Progression-free survival of SMAD4-WT (blue) and SMAD4 mutant or SMAD4 null (red) TCGA pancreatic cancer primary resected tumors (p = 0.0179); see . (F) Quantification of tumor area (percentage of tumor area per lung lobe) in lungs of immunodeficient mice i.v. inoculated with SMAD4 isogenic pancreatic cancer cells; each lobe is presented as an independent data point. n = 3 or 4 mice (t test, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001); see . (G) Longitudinal quantification of BLI of mice tail vein inoculated with LacZ- or SMAD4-expressing HPAC cells with BLI (red indicates HPAC SMAD4; blue indicates HPAC LacZ; t test, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; n = 7 for each cohort). (H) Representative immunofluorescent images of mice i.v. inoculated with LacZ- or SMAD4-expressing HPAC cells at 12 weeks following inoculation. SMAD4 cells are stained in yellow (scale bars, 100 μm).

Article Snippet: PANC1 , Broad Institute , Project Achilles.

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Imaging, Standard Deviation, Injection, Mutagenesis, Staining

Journal: Cell reports

Article Title: SMAD4 represses FOSL1 expression and pancreatic cancer metastatic colonization

doi: 10.1016/j.celrep.2021.109443

Figure Lengend Snippet: (A) Representative bioluminescent signal images of mice (n = 10 for each cohort) 12 weeks after tail vein inoculation with HPAC cell lines with sh-scramble, sh-FOSL1 hairpin 1, or sh-FOSL1 hairpin 2. (B) Immunoblot showing reduced FOSL1 expression in xenograft tumors formed from SMAD4-expressing HPAC cells compared to those formed from LacZ-expressing HPAC cells. (C) qRT-PCR of FOSL1 in PANC1 cells with SMAD4 gene deletion treated 24 h with TGF-β. Mean of three biological replicates is shown (t test, **p < 0.01; standard deviation is reported in error bars) (D) ChIP qPCR on FOSL1 promoter, enhancer, and gene body in LacZ- and SMAD4-expressing HPAC cells treated with TGF-β. Mean of four (for ChIP/SMAD4) or five (for ChIP/H3K27Ac) biological replicates is shown (standard deviation is reported in error bars). (E) ChIP qPCR on FOSL1 promoter, enhancer, and gene body in PANC1 cells with SMAD4 gene deletion treated with TGF-β. Mean of three (for ChIP/SMAD4) or six (for ChIP/H3K27Ac) biological replicates is shown (standard deviation is reported in error bars).

Article Snippet: PANC1 , Broad Institute , Project Achilles.

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Standard Deviation, ChIP-qPCR

Journal: Cell reports

Article Title: SMAD4 represses FOSL1 expression and pancreatic cancer metastatic colonization

doi: 10.1016/j.celrep.2021.109443

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: PANC1 , Broad Institute , Project Achilles.

Techniques: Virus, Recombinant, Reverse Transcription, SYBR Green Assay, Software