Journal: iScience

Article Title: Compartmentalization of casein kinase 1 γ CSNK1G controls the intracellular trafficking of ceramide

doi: 10.1016/j.isci.2022.104624

Figure Lengend Snippet:

Article Snippet: anti-GAPDH , Fujifilm Wako Pure Chemical Corporation , Cat#: 016-25523; RRID: AB_2814991.

Techniques: Transduction, Recombinant, Protease Inhibitor, Software

Journal: American Journal of Cancer Research

Article Title: Stat5b inhibition blocks proliferation and tumorigenicity of glioblastoma stem cells derived from a de novo murine brain cancer model

doi:

Figure Lengend Snippet: Lgr5 and Stat5b expression are induced by hypoxia in GSCs. A. Representative images of immunohistochemical staining for Hif2α, Lgr5, and Stat5b in human GBM tissue. B. Stat5b mRNA levels were analyzed by qRT-PCR in GSCs cultured in hypoxic conditions for 2 days (0.5% O2; n=3; ***P<0.001). C. Western blot analysis of phospho-Stat5 and Stat5b in GSCs cultured in hypoxic conditions (0.5% O2 for 4 days, 5% O2 for 3 days). β-tubulin is shown as a loading control. D. Western blot analysis of Hif2α and Stat5b in GSCs after treatment with control-shRNA (Control) or Hif2α-shRNA (Hif2α-KD) for 4 days. GAPDH is shown as a loading control. E. Lgr5 mRNA expression levels were analyzed by qRT-PCR in GSCs after 2 days in hypoxic conditions (0.5% O2; n=3; **P<0.01). F. Lgr5 expression levels were analyzed by a flow cytometry in the GSCs after 2 days in hypoxic conditions (0.5% O2).

Article Snippet: The following antibodies were used: Stat5b (1:1000; ab178941, Abcam), Lgr5 (1:500; bs-1117R; Bioss, Woburn, MA, USA), Hif2α (1:500; NB 100-122; Novus Biologicals, Centennial, CO, USA), β-tubulin (1:200; T4026, Sigma-Aldrich), GAPDH (1:1000; 016-25523, Wako), Oct4 (1:1000; ab19875, Abcam), Survivin (1:1000; ab182132, Abcam), Cyclin E2 (1:500; ab32103, Abcam), Phospho-Rb (1:1000; #8516, Cell Signaling Technology [CST], Danvers, MA, USA), Phospho-Stat5 (1:1000; ab32364, Abcam), Caspase3 (1:1000; #9665, CST), PARP (1:1000; #9542, CST), β-Catenin (1:1000; #9587, CST).

Techniques: Expressing, Immunohistochemical staining, Staining, Quantitative RT-PCR, Cell Culture, Western Blot, shRNA, Flow Cytometry

Journal: American Journal of Cancer Research

Article Title: Stat5b inhibition blocks proliferation and tumorigenicity of glioblastoma stem cells derived from a de novo murine brain cancer model

doi:

Figure Lengend Snippet: Stat5b expression in GSCs regulated by Wnt pathway. A. GSCs were treated with control-shRNA (Control) or Lgr5-shRNA (Lgr5-KD) for 5 days, and then Lgr5, β-Catenin, and Stat5b were analyzed by western blot. GAPDH is shown as a loading control. B. GSCs were treated with ICG-001 for 48 h, and the number of viable cells was assessed using a trypan blue dye exclusion test (n=4; ***P<0.001). C, D. The percentage of BrdU-incorporated cells treated with DMSO (Control) or ICG-001 (5 μM) for 48 h was determined by flow cytometry (n=3; *P<0.05). E, F. Flow cytometric analysis of Annexin V/PI staining for the detection of apoptosis in GSCs treated with DMSO (Control) or ICG-001 (5 μM) for 48 h (n=6; **P<0.001). G. Western blot analyses of Stat5b, OCT4, and Survivin in independent GSC lines treated with ICG-001 for 48 h. GAPDH is shown as a loading control.

Article Snippet: The following antibodies were used: Stat5b (1:1000; ab178941, Abcam), Lgr5 (1:500; bs-1117R; Bioss, Woburn, MA, USA), Hif2α (1:500; NB 100-122; Novus Biologicals, Centennial, CO, USA), β-tubulin (1:200; T4026, Sigma-Aldrich), GAPDH (1:1000; 016-25523, Wako), Oct4 (1:1000; ab19875, Abcam), Survivin (1:1000; ab182132, Abcam), Cyclin E2 (1:500; ab32103, Abcam), Phospho-Rb (1:1000; #8516, Cell Signaling Technology [CST], Danvers, MA, USA), Phospho-Stat5 (1:1000; ab32364, Abcam), Caspase3 (1:1000; #9665, CST), PARP (1:1000; #9542, CST), β-Catenin (1:1000; #9587, CST).

Techniques: Expressing, shRNA, Western Blot, Flow Cytometry, Staining