anti-stat5 cell signaling 9363  (Cell Signaling Technology Inc)

Journal: Scientific Reports

Article Title: GRAM domain-containing protein 1A (GRAMD1A) promotes the expansion of hepatocellular carcinoma stem cell and hepatocellular carcinoma growth through STAT5

doi: 10.1038/srep31963

Figure Lengend Snippet: ( a) Analysis of the correlation between GRAMD1A levels and STAT5 levels using GSEA. ( b) Quantitative real-time PCR assay for the mRNA levels of STAT5, CyclinD1, Bcl-2, c-jun and c-Myc after GRAMD1A overexpression or knockdown. ( c) Luciferase activity assay was used to determine whether GRAMD1A could regulate the transcriptional activity of STAT5. Data are expressed as mean ± SEM. *p < 0.05.

Article Snippet: The following antibodies were used: GRAMD1A (Sigma, HPA008852), caspase-3 (Cell Signaling Technology, #9662), poly(ADP-ribose) polymerase (PARP) (Cell Signaling Technology, #9542), BCL-XL (Cell Signaling Technology, #2762), CD133 (Miltenyi Biotec, 130-092-395), CD90 (Abcam, ab133350), STAT5 (Cell Signaling Technology, #9363), β-actin (Abcam, ab8226) and GAPDH (Abcam, ab8245).

Techniques: Real-time Polymerase Chain Reaction, Over Expression, Knockdown, Luciferase, Activity Assay

Journal: Scientific Reports

Article Title: GRAM domain-containing protein 1A (GRAMD1A) promotes the expansion of hepatocellular carcinoma stem cell and hepatocellular carcinoma growth through STAT5

doi: 10.1038/srep31963

Figure Lengend Snippet: ( a) Hepatosphere formation assay for the effect of STAT5 downregulation on self-renewal of HCC stem cells in Huh-7 and HepG2 cells with GRAMD1A overexpression. Representative micrographs ( left ); quantification of hepatosphere number ( right ). ( b) SP analysis demonstrated the role of STAT5 downregulation in self-renewal of HCC stem cells in Huh7 and HepG2 with GRAMD1A overexpression. ( c) TUNEL assay for the effect of STAT5 downregulation on resistance to Doxorubicin in Huh7 and HepG2 with overexpression. Representative micrographs ( left ); quantification of TUNEL positive cell number ( right ). DAPI was used to stain nucleus. ( d) Western blot assay for caspase 3 activity, PARP cleavage and BCL-X L levels after STAT5 downregulation in Huh-7 and HepG2 with GRAMD1A overexpression. ( e) Soft agar growth ability assay for the role of STAT5 downregulation in the anchorage-independent cell growth of Huh7 and HepG2 with GRAMD1A overexpression. Representative micrographs ( left ); quantification of colony number with diameter greater than 0.1 mm ( right ). Data are expressed as mean ± SEM. *p < 0.05.

Article Snippet: The following antibodies were used: GRAMD1A (Sigma, HPA008852), caspase-3 (Cell Signaling Technology, #9662), poly(ADP-ribose) polymerase (PARP) (Cell Signaling Technology, #9542), BCL-XL (Cell Signaling Technology, #2762), CD133 (Miltenyi Biotec, 130-092-395), CD90 (Abcam, ab133350), STAT5 (Cell Signaling Technology, #9363), β-actin (Abcam, ab8226) and GAPDH (Abcam, ab8245).

Techniques: Tube Formation Assay, Over Expression, TUNEL Assay, Staining, Western Blot, Activity Assay