Journal: Neoplasia (New York, N.Y.)

Article Title: Abscisic acid regulates dormancy of prostate cancer disseminated tumor cells in the bone marrow

doi: 10.1016/j.neo.2020.11.009

Figure Lengend Snippet: ABA induces cellular dormancy through PPARγ receptor signaling in PCa cells. (A) Basal levels of mRNA expression of the receptor of ABA, PPARγ in PCa cell lines as quantified by real-time PCR. (B) Basal levels of protein expression of the receptor of ABA, PPARγ in PCa cell lines as quantified by Western blot. (C) mRNA expression of PPARγ in G 0 , G 1 , and S/G 2 /M cell cycle phase of PC3 VC cells. (D) Verification of PPARγ mRNA expression in PPARγ silencing PC3 VC as quantified by real-time PCR. (E) Proliferation assays in PC3 VC -Control or PC3 VC -shPPARγ cells were performed in 1% FBS culture condition with ABA treatment. (F)% of G 0, G 1, or S/G 2 /M cell cycle phase in PC3 VC -Control or PC3 VC -shPPARγ cells following vehicle or ABA (50 µM) treatment in 1% FBS culture conditions at 72 h as quantified by FACS analyses. Data in C–F are representative of mean with SD (Student's t test). (G) Activation of down-stream effector, p70S6K of mTOR signaling in PC3 VC -Control or PC3 VC -shPPARγ cells following vehicle or ABA (50 µM) treatment as quantified by Western blot.

Article Snippet: Lentiviral particles with PPARγ human shRNA or negative control (cat no. TL320459V , Origene, Rockville, MD) were infected into PC3 VC cells in the presence of 5 μg/mL polybrene for 48 h. Infected cells were selected for 7 d in media containing 1 μg/mL Puromycin and analyzed by real-time PCR or ELISA.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control, Activation Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: Abscisic acid regulates dormancy of prostate cancer disseminated tumor cells in the bone marrow

doi: 10.1016/j.neo.2020.11.009

Figure Lengend Snippet: ABA and PPARγ signaling pathway induces PCa cellular dormancy in the bone marrow microenvironment. (A) ABA production was identified in culture media of MC3TC-E1 cells at 72 h as quantified by ELISA (cat. LS- F4483-1, Life Span Biosciences). ABA production was normalized to total protein. (B) % of G 0, G 1, or S/G 2 /M cell cycle phase when PC3 VC -Control or PC3 VC -shPPARγ cells are co-cultured with murine osteoblasts (MC3T3-E1 cells) with or without ABA (50 µM) treatment. Live cells (cat. NBP2-31156, DAPI, NOVUS) were negatively gated for anti-mouse H-2kd (cat no. 116622, PE/Cy7, BioLegend), which were then positively gated for (human) HLA-A,B,C (cat no. 311426, APC/Cy7, BioLegend). After these gates were applied, the cells were plotted on the Venus-Cherry spectrum. Cell cycle phase was determined using FACS analyses. (C) Diagram of the experimental procedures for in vivo animal model. PC3 VC -Control or PC3 VC -shPPARγ cells (2 × 10 5 cells) were suspended in 30μl of PBS and injected into 5- to 7-wk-old male CB.17. SCID mice by i.t. injection. After PCa cell injection, vehicle or ABA (20 mg/kg) treatment for 8 times (twice daily) was followed by i.p. injection. At 4 d, mice were sacrificed, and the tibiae which PCa cells were injected were collected for analyzing cell cycle phase of PC3 VC cells by FACS analyses. Antibody staining were applied as same as in coculture study in A. (D) Quantification of % of G 0, G 1, or S/G 2 /M cell cycle phase from 5C ( n = 4/group). Data in A, B and D are representative of mean with SD (Student's t test).

Article Snippet: Lentiviral particles with PPARγ human shRNA or negative control (cat no. TL320459V , Origene, Rockville, MD) were infected into PC3 VC cells in the presence of 5 μg/mL polybrene for 48 h. Infected cells were selected for 7 d in media containing 1 μg/mL Puromycin and analyzed by real-time PCR or ELISA.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Cell Culture, In Vivo, Animal Model, Injection, Staining

Journal: Cell reports

Article Title: Mapping Distinct Bone Marrow Niche Populations and Their Differentiation Paths

doi: 10.1016/j.celrep.2019.06.031

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: shRNA - TF510219 (Pparg) , Origene , 4 unique shRNA.

Techniques: Recombinant, shRNA, Construct, Plasmid Preparation, Sequencing, Software

Journal: Oncotarget

Article Title: Alpha-Tocopherol prevents esophageal squamous cell carcinoma by modulating PPARγ-Akt signaling pathway at the early stage of carcinogenesis.

doi: 10.18632/oncotarget.21437

Figure Lengend Snippet: Figure 6: PPARγ played an important role in the suppression of Akt signaling by α-Tocopherol. (A) Western blotting of PPARγ and PTEN in rat esophagus. Two individual pooled samples from 12 rat of each group were analyzed. (B) Immunostaining of PPARγ and PTEN in rat esophagus. (C) Western blotting of PPARγ and PTEN in Het-1A cells. Het-1A cells were treated with NMBA (50, 100 μM) with or without α-Tocopherol (25, 50, 100 μM) for 24h. (D) Western blotting of PPARγ and Akt pathway in Het-1A cells. Het- 1A cells were treated with Rosiglitazone (RGZ, 20 μM) or GW9662 (GW, 20 μM) for 24h. (E) Proteomic array analysis on Akt signaling pathway. Het-1A cells were transfected with si-PPARG and then treated with α-Tocopherol (50 μM) or Rosiglitazone (RGZ, 20 μM) for 24h. Data are shown as mean ± standard deviation. #, p<0.05 compared with the control group; *, p<0.05 compared with the si-PPARG group.

Article Snippet: The Trilencer-27 human PPARG siRNA (siPPARG) and Trilencer-27 universal scrambled negative control siRNA (si-CT) were purchased from OriGene Technologies, Inc. (Rockville, MD, USA).

Techniques: Western Blot, Immunostaining, Transfection, Standard Deviation, Control