Journal: bioRxiv

Article Title: Stanniocalcin 1a is a Ca 2+ -regulated switch controlling epithelial cell quiescence-proliferation balance and Ca 2+ uptake

doi: 10.1101/2020.09.09.290114

Figure Lengend Snippet: ( A-B ) Elevated NaR cell proliferation in stc1a -/- fish. Progenies of stc1a (+17) +/- intercrosses were raised in the E3 embryo medium to 5 dpf. NaR cells were detected by in situ hybridization using an igfbp5a cRNA probe. After NaR cells were visualized and quantified in each fish, fish was genotyped individually. Representative images are shown in ( A ) and quantified data in ( B ). Scale bar = 0.2 mm. n = 33-70 larvae/group. (C ) Increased IGF1 receptor-Akt signaling in stc1a -/- fish. Progenies of stc1a (+17) +/- ; Tg (igfbp5a: GFP) intercrosses were raised in E3 embryo medium to 3 dpf and treated with DMSO or 0.3 μM BMS-754807 (BMS). Two days later, fish were fixed and phospho-Akt positive cells were detected by immunostaining. These fish were genotyped individually afterwards. n = 14-47 larvae/group. ( DE) Progeny of stc1a (+17) +/- ; Tg (igfbp5a: GFP) intercrosses were raised in E3 embryo medium and transfer to normal [Ca 2+ ] embryo medium containing DMSO, 0.3 μM BMS-754807 (BMS), 0.06 μM Wortmannin (Wort), 8 μM MK2206 (MK), 5 μM Rapamycin (Rapa), or 10 μM U0126 at 3 dpf. Two days later, NaR cells were quantified. They were genotyped individually afterwards. Representative images ( D ) and quantified data are shown ( E ). n = 7-35 larvae/group. Scale bar = 0.2 mm.

Article Snippet: Alizarin Red S, ZnCl 2 , batimastat and GdCl 3 were purchased from Sigma (St. Louis, MO, USA), MK2206 from ChemieTek (Indianapolis, IN) and Rapamycin from Calbiochem (Gibbstown, NJ).

Techniques: In Situ Hybridization, Immunostaining

Journal: bioRxiv

Article Title: The Hexosamine Biosynthetic Pathway alters the cytoskeleton to modulate cell proliferation and migration in metastatic prostate cancer

doi: 10.1101/2024.10.14.618283

Figure Lengend Snippet: (A-C) Control and GNPNAT1 KO 22Rv1 cell lysate samples were Western blot-analyzed for the levels of AKT and its phosphorylated form p-AKT (A) as well as AKT downstream signaling pathways including components of the mTOR pathway (B), and the PKC pathway (C). (D-E) Microscopic images (D) and cell proliferation assay (E) after treating control and GNPNAT1 KO with the AKT inhibitor, MK2206 after 48 h. Scale bars, 200 µm. F) Cell proliferation assay after treatment with UDP-GlcNAc after 48 h. (G) Cell proliferation assay after pre-treatment with 10 µM MK2206 followed by co-treatment with 10 µM MK2206 and 30 µM UDP-GlcNAc or UDP-GlcNAc alone. (H) Schematic diagram of the possible mechanism for increased cell proliferation in GNPNAT1 KO cells, via the alteration of AKT and its downstream signaling pathways.

Article Snippet: MK2206 was purchased from AdooQ Bioscience (Irvine, CA) and UDP-GlcNAc from Millipore sigma (Burlington, MA).

Techniques: Control, Western Blot, Proliferation Assay

Journal: International Journal of Biological Sciences

Article Title: The HDAC Inhibitor Quisinostat (JNJ-26481585) Supresses Hepatocellular Carcinoma alone and Synergistically in Combination with Sorafenib by G0/G1 phase arrest and Apoptosis induction

doi: 10.7150/ijbs.27661

Figure Lengend Snippet: PI3K/AKT and JNK/c-Jun pathway facilitated by Quisinostat were required for cell cycle arrest and cell apoptosis. (A) HCCLM3 and SMMC-7721 cells were treated with quisinostat (12.5, 25.0 and 50.0nM) for 48 h. Western blot analysis was used to evaluate expressions of PI3K-p110, PI3K-p85, phosphorylation of AKT 473 , JNK, phosphorylation of JNK and c-Jun. Data, mean ± SD (n = 3); *, P < 0.05; **, P < 0.01 compared with DMSO group. (B) Cells were preincubated for 5 h with or without MK2206 2HCL(5μM), followed by culturing with 25nM quisinostat for 48 h, then conducting the cell cycle analysis and (C) Expression levels of phosphorylation of AKT 473 and p21 proteins were detected by Western Blotting. Data were shown as mean ± SD. n = 3; *, P < 0.05; **, P < 0.01; ***, P < 0.001 compared with the DMSO group. (D) Cells were pretreated with or without 40μM Z-VAD-FMK for 2 h, followed by culturing with 25nM quisinostat for 48 h to conduct apoptosis analysis, (E) cell proliferation analysis and (F) Expression levels of cle-Caspase9, cle-Caspase3, PARP, and cle-PARP proteins were detected by Western Blotting. Data were shown as mean ± SD. n = 3; *P <0.05; **P < 0.01; ***P < 0.001 compared with Quisinostat group. (G) Cells were pretreated with SP600125 (6μM) for 5 h followed by culturing with 25nM quisinostat for 48 h to conduct apoptosis analysis, (H) cell proliferation analysis and (I) Expression levels of phosphorylation of JNK, cle-Caspase3 and cle-PARP proteins were detected by Western Blotting. Data were shown as mean ± SD. n = 3; *P <0.05; **P < 0.01; ***P < 0.001 compared with Quisinostat group.

Article Snippet: Sorafenib, quisinostat (JNJ-26481585), JNK inhibitor SP600125, AKT inhibitor MK2206 2HCL and Caspase inhibitor Z-VAD-FMK were from Selleckchem (Houston, Texas, USA).

Techniques: Western Blot, Phospho-proteomics, Cell Cycle Assay, Expressing