protoscript ii rt pcr kit  (New England Biolabs)

Journal: Cancer Science

Article Title: Estrogen receptor variant ER ‐α36 promotes tamoxifen agonist activity in glioblastoma cells

doi: 10.1111/cas.13868

Figure Lengend Snippet: Tamoxifen ( TAM ) induced glioma cell autophagy. A, U87‐ MG cells were treated with 5 μmol/L TAM for different time periods. Mitochondrial membrane potentia was analyzed using flow cytometry. B, Western blot analysis of the expression of P53 and Bcl‐2. C, Cells were treated with 5 μmol/L TAM for different time periods, and then stained with acridine orange and examined under a fluorescence microscope. D, Western blot analysis of the accumulation of LC 3‐II in U87‐ MG cells that were treated with TAM (5 μmol/L) for different time period. E, p‐Akt, Akt, p‐ mTOR and mTOR protein levels were estimated by western blot analysis. F, RT ‐ PCR examination of autophagy pathway in mRNA expression. Beclin‐1, ATG ‐5, ATG ‐12, P62 (n = 5, * P < 0.05, ** P < .01 vs control). mTOR , mammalian target of rapamycin

Article Snippet: Total RNA (1 μg) was reversely transcribed using the ProtoScript II RT‐PCR kit (Takara, Dalian, China).

Techniques: Flow Cytometry, Western Blot, Expressing, Staining, Fluorescence, Microscopy, Reverse Transcription Polymerase Chain Reaction

Journal: Cancer Science

Article Title: Estrogen receptor variant ER ‐α36 promotes tamoxifen agonist activity in glioblastoma cells

doi: 10.1111/cas.13868

Figure Lengend Snippet: Overexpression of ER ‐α36 in U251 cells leads to decreased sensitivity to tamoxifen ( TAM ) and enhanced autophagy. U251 cells overexpressed ER ‐α36. A, RT ‐ PCR examination of ER ‐α36 expression. B, MTT analysis of cell viability of glioma cells. C, Western blot analysis of the accumulation of LC 3‐II (n = 4, ** P < .01, * P < .05 vs control). ER , estrogen receptor

Article Snippet: Total RNA (1 μg) was reversely transcribed using the ProtoScript II RT‐PCR kit (Takara, Dalian, China).

Techniques: Over Expression, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot

Journal: Cancer Science

Article Title: Estrogen receptor variant ER ‐α36 promotes tamoxifen agonist activity in glioblastoma cells

doi: 10.1111/cas.13868

Figure Lengend Snippet: U251/ TAM shows increased levels of ER ‐α36, together with increased cell autophagy. A, RT‐PCR examination of ER‐α36 expression in U251 and U251/TAM. B, Knockdown of ER‐α36 by shRNA method. RT‐PCR examination of ER‐α36 expression in U251/TAM‐V and U251/TAM‐siRNA. C, MTT analysis of cell viability of glioma cells. D, Western blot analysis of accumulated LC 3‐II (n = 4, ** P < .01 vs control). ER , estrogen receptor; TAM, tamoxifen

Article Snippet: Total RNA (1 μg) was reversely transcribed using the ProtoScript II RT‐PCR kit (Takara, Dalian, China).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, shRNA, Western Blot