Journal: Integrative zoology

Article Title: cba-miR-222-3p involved in photoperiod-induced apoptosis in testes of striped hamsters by targeting TRAF7.

doi: 10.1111/1749-4877.12918

Figure Lengend Snippet: Figure 3 Validation of cba-miR-222-3p targeting TRAF7 and TRAF7 expression in the testes of striped hamsters. (a) Sequences and peak maps of cba-miR-222-3p, TRAF7-WT, and TRAF7-MT. (b) Relative luciferase activity detected by Dual-Luciferase Reporter Assay. (c) Immunohistochemistry (IHC, tissue microarray [TMA]) of TRAF7 in testes. (d) Integrated density of TRAF7 detected by IHC (TMA; n = 4). (e) Protein expression levels of TRAF7 in the testes detected by western blot (n = 4). (f) Pearson correlation analysis of cba-miR-222-3p and TRAF7. LD, long daylength; MD, moderate daylength; SD, short daylength; ∗, P < 0.05; ∗∗, P < 0.01.

Article Snippet: After performing electrophoresis, the proteins were transferred to PVDF membranes, which were then incubated with TRAF7 antibodies (11780-1-AP, Proteintech, China, RRID:AB_2877793) at a 1:1000 dilution and β-actin antibodies (20536-1-AP, Proteintech, China, RRID:AB_10700003) at a 1:1000 dilution, and subsequently incubated with IRDye 800 CW goat anti-rabbit secondary antibodies (31 460, Thermo Fisher, USA).

Techniques: Biomarker Discovery, Expressing, Luciferase, Activity Assay, Reporter Assay, Immunohistochemistry, Microarray, Western Blot

Journal: Integrative zoology

Article Title: cba-miR-222-3p involved in photoperiod-induced apoptosis in testes of striped hamsters by targeting TRAF7.

doi: 10.1111/1749-4877.12918

Figure Lengend Snippet: Figure 4 Expression of cba-miR-222-3p and TRAF7 and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) analysis after in vivo injection in the testes of striped hamsters. (a) Schematic diagram of in vivo injection experiment of miRNA mimics. (b) Fluorescence in situ hybridization (FISH, tissue microarray [TMA]) of cba-miR-222-3p in testes after in vivo injection. (c) Fluorescence intensity of cba-miR-222-3p detected by FISH (TMA; n = 4). (d) Immunohistochemistry (IHC, TMA) of TRAF7 in testes after in vivo injection. (e) Integrated density of TRAF7 detected by IHC (TMA; n = 4). (f) TUNEL (TMA) staining of the testes after in vivo injection. (g) The proportion of TUNEL (TMA) staining with apoptotic activity in the testes (n = 4). (h) Relative expression of MEKK3 (n = 6). (i) Relative expression of p38 (n = 6). (j) Relative expression of p53 (n = 6). AG, agomir injection; NC, agomir negative control injection; DAPI, 4′6′-diamidino-2-phenylindole. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001.

Article Snippet: After performing electrophoresis, the proteins were transferred to PVDF membranes, which were then incubated with TRAF7 antibodies (11780-1-AP, Proteintech, China, RRID:AB_2877793) at a 1:1000 dilution and β-actin antibodies (20536-1-AP, Proteintech, China, RRID:AB_10700003) at a 1:1000 dilution, and subsequently incubated with IRDye 800 CW goat anti-rabbit secondary antibodies (31 460, Thermo Fisher, USA).

Techniques: Expressing, TUNEL Assay, In Vivo, Injection, Fluorescence, In Situ Hybridization, Microarray, Immunohistochemistry, Staining, Activity Assay, Negative Control

Journal: PLoS Genetics

Article Title: A SHH-FOXF1-BMP4 signaling axis regulating growth and differentiation of epithelial and mesenchymal tissues in ureter development

doi: 10.1371/journal.pgen.1006951

Figure Lengend Snippet: (A) Pie-chart summarizing the results from the microarray analysis of E12.5 ureters explanted and treated with DMSO or 10 μM cyclopamine for 18 h filtered with an intensity (Int) threshold of 150 and a fold change (FC) cut-off of 2.0. (B) Table of the downregulated transcripts. Shown are average intensities of transcripts in control and cyclopamine treated ureters and average fold changes (FC) of RNA intensities between the pools in two independent experiments. (C) In situ hybridization analysis of expression of microarray candidates on proximal ureter sections of control, Tbx18 cre/+ ; Smo fl/fl ( Smo LOF ) and Tbx18 cre/+ ; R26 mTmG/SmoM2 ( Smo GOF ) ureters at E12.5 and E14.5.

Article Snippet: For pharmacological manipulation of SHH signaling cyclopamine (Selleckchem) and purmorphamine (Millipore) were used at a final concentration of 10 μM and 2 μM, respectively.

Techniques: Microarray, Control, In Situ Hybridization, Expressing

Journal: PLoS Genetics

Article Title: A SHH-FOXF1-BMP4 signaling axis regulating growth and differentiation of epithelial and mesenchymal tissues in ureter development

doi: 10.1371/journal.pgen.1006951

Figure Lengend Snippet: (A-U) Ureters were explanted from E12.5 wildtype, Axin2 creERT2/+ ; Hprt Foxf1/y , Tbx18 cre/+ ; Hprt Foxf1DN/y or Tbx18 cre/+ ; R26 mTmG/+ embryos and cultured for 6 d in the presence or absence of 10 μM cyclopamine, 100 ng/μl BMP4, 10 μg/ml NOGGIN, 2 μM Purmorphamine or solvent as indicated. Whole explants were documented by epifluorescence analysis (Q), or were sectioned and proximal regions analyzed by Haematoxylin and Eosin staining (A,E,I,M,R), by immunofluorescence (B-D,F-H,J-L,N-P,S-U) for the SMC marker ACTA2 together with the epithelial marker CDH1 (B,F,J,N,S), for the SMC marker TAGLN without (C,G,K,O) or with the lineage marker GFP (T), and for the urothelial markers ΔNP63/UPK1B (D,H,L,P,U).

Article Snippet: For pharmacological manipulation of SHH signaling cyclopamine (Selleckchem) and purmorphamine (Millipore) were used at a final concentration of 10 μM and 2 μM, respectively.

Techniques: Cell Culture, Solvent, Staining, Immunofluorescence, Marker