Journal: Journal of pharmacological sciences

Article Title: Lymphangiogenesis induced by vascular endothelial growth factor receptor 1 signaling contributes to the progression of endometriosis in mice.

doi: 10.1016/j.jphs.2020.05.003

Figure Lengend Snippet: Fig. 3. Accumulation of macrophages and fibroblasts in endometrial implant tissues. (A) The levels of the mRNAs encoding the lymphangiogenic growth factors VEGF-C and VEGF-D in endometrial tissues from WT/WT and TK-/-/TK-/- mice. Data are expressed as the mean ± SEM (n ¼ 5e6 mice per group). *P < 0.05. (B) Double immunostaining of VEGF-C (green) or VEGF-D (green), and CD11b (red) or S100A4 (red) in endometrial tissues from WT/WT mice at Day 14. Scale bars, 50 mm. (C) Left: Immunostaining of CD11b (green) in endometrial tissues from WT/WT and TK-/-/TK-/- mice at Day 14. Scale bars, 100 mm. Right: The numbers of CD11bþ cells in endometrial tissues from WT/WT and TK-/-/TK-/-

Article Snippet: The sections were incubated with the following primary antibodies at 4 C overnight: a rabbit anti-mouse VEGFR1 polyclonal antibody (1:200; Abcam), a goat anti-mouse VEGFR1 polyclonal antibody (1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), a rabbit anti-LYVE-1 polyclonal antibody (1:200; Abcam), a goat anti-LYVE-1 polyclonal antibody (1:200; R&D Systems Inc., Minneapolis, MN, USA), a rabbit anti-VEGF-C polyclonal antibody (1:200; Abcam), a goat anti-VEGFD polyclonal antibody (1:40; R&D Systems Inc.), a rat anti-mouse CD11b monoclonal antibody (1:200; BD Biosciences, San Jose, CA, USA), a rabbit anti-mouse S100A4 polyclonal antibody (1:200; Abcam), or a goat anti-mouse S100A4 polyclonal antibody (1:100; OriGene Technologies, Rockville, MD, USA).

Techniques: Double Immunostaining, Immunostaining

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Exosomal microRNA ‐15a from ACHN cells aggravates clear cell renal cell carcinoma via the BTG2 / PI3K / AKT axis

doi: 10.1002/kjm2.12428

Figure Lengend Snippet: miR‐15a targets BTG2 in ccRCC cells. (A) Targeting relation between miR‐15a and BTG2 verified by dual‐luciferase reporter gene assay (* p < 0.05; compared using two‐way ANOVA). (B) BTG2 expression in UMRC‐2 cells treated with exosomes determined by RT‐qPCR (* p < 0.05; compared using unpaired t test). (C) BTG2 expression in ccRCC tissues and adjacent normal tissues detected by RT‐qPCR (* p < 0.05; compared using paired t test). (D) Correlation analysis between miR‐15a and BTG2 in ccRCC tissues. (E) BTG2 expression in ccRCC cells and HK‐2 cells detected by RT‐qPCR (* p < 0.05; compared using one‐way ANOVA). (F) BTG2 expression quantified by RT‐qPCR following altered expression of miR‐15a (* p < 0.05, ** p < 0.01; compared using one‐way ANOVA). (G) Transfection efficiency of BTG2‐OE in cells verified by RT‐qPCR (* p < 0.05; compared using unpaired t test). (H) Effects of BTG2 on ccRCC cell proliferation determined by colony formation assay (* p < 0.05; compared using unpaired t test)

Article Snippet: The first five nucleotides complementary to the seed region of miR‐15a were mutated by site‐directed mutagenesis, and the full‐length BTG2 cDNA (OriGen Biomedical, Austin, TX) without the 3′‐UTR was subcloned into the eukaryotic expression vector PCMM‐ENTRY.

Techniques: Luciferase, Reporter Gene Assay, Expressing, Quantitative RT-PCR, Transfection, Colony Assay

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Exosomal microRNA ‐15a from ACHN cells aggravates clear cell renal cell carcinoma via the BTG2 / PI3K / AKT axis

doi: 10.1002/kjm2.12428

Figure Lengend Snippet: BTG2 can offset the effects of miR‐15a on ccRCC cells. (A) Establishment of cells overexpressing both miR‐15a and BTG2 verified by RT‐qPCR (* p < 0.05; compared using unpaired t test). (B) Cell proliferation detected by colony formation assay (* p < 0.05; compared using unpaired t test). (C) Cell migration examined by transwell assay (* p < 0.05; compared using unpaired t test). (D) Cell invasion evaluated by transwell assay (* p < 0.05; compared using unpaired t test). (E) Cell apoptosis determined by flow cytometry (* p < 0.05; compared using unpaired t test). (F) EMT activity of cells shown by detecting the florescence intensity of E‐cadherin

Article Snippet: The first five nucleotides complementary to the seed region of miR‐15a were mutated by site‐directed mutagenesis, and the full‐length BTG2 cDNA (OriGen Biomedical, Austin, TX) without the 3′‐UTR was subcloned into the eukaryotic expression vector PCMM‐ENTRY.

Techniques: Quantitative RT-PCR, Colony Assay, Migration, Transwell Assay, Flow Cytometry, Activity Assay

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Exosomal microRNA ‐15a from ACHN cells aggravates clear cell renal cell carcinoma via the BTG2 / PI3K / AKT axis

doi: 10.1002/kjm2.12428

Figure Lengend Snippet: miR‐15a‐mediated BTG2 inhibition increases activity of the PI3K/AKT pathway. Changes of PI3K/AKT pathway in exosomes‐treated cells or cells overexpressing BTG2 or/and miR‐15a detected by western blot analysis

Article Snippet: The first five nucleotides complementary to the seed region of miR‐15a were mutated by site‐directed mutagenesis, and the full‐length BTG2 cDNA (OriGen Biomedical, Austin, TX) without the 3′‐UTR was subcloned into the eukaryotic expression vector PCMM‐ENTRY.

Techniques: Inhibition, Activity Assay, Western Blot