shrictor  (Addgene inc)

Journal: Oncology Research

Article Title: mTORC2 promotes pancreatic cancer progression and parp inhibitor resistance

doi: 10.32604/or.2023.029309

Figure Lengend Snippet: The mTORC2 obligate component Rictor is overexpressed and associated with poor survival in PDAC patients. (A) Analysis of Rictor mRNA levels in pancreatic adenocarcinoma (PAAD) tumor (T) tissues (n = 179) and normal pancreas (N) tissues (n = 171) through The Cancer Genome Atlas (TCGA) database. (B) IHC analysis of Rictor protein expression in pancreatic ductal adenocarcinoma (PDAC) tissues and normal pancreas tissues. Representative images of IHC staining (left) and quantification of the IHC score (right) are shown. Data are presented as the mean ± standard deviation (SD), n = 10. *** p < 0.001 according to 2-tailed t test. (C) Analysis of Rictor protein level in different stage of PDAC tumors. * p < 0.05 by 2-tailed t test. (D) Kaplan‒Meier survival analysis of Rictor expression in PDAC patients (n = 178) from the TCGA database.

Article Snippet: Anti-β-actin antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). pDR-GFP (#17617), pCBASceI (#26477) and Rictor shRNA plasmids (#1853 and #1854) were obtained from Addgene (Watertown, MA, USA).

Techniques: Expressing, Immunohistochemistry, Standard Deviation

Journal: Oncology Research

Article Title: mTORC2 promotes pancreatic cancer progression and parp inhibitor resistance

doi: 10.32604/or.2023.029309

Figure Lengend Snippet: Knockdown of Rictor suppresses PDAC cell growth and invasion. (A) Western blot analysis of Rictor levels in PANC-1 or MIA PaCa-2 cells expressing Rictor shRNA. (B) Cell growth analysis of cells expressing Rictor shRNA. (C) Transwell invasion assay of ANC-1 or MIA PaCa-2 cells stably transfected with control (Ctrl) or Rictor shRNA. Representative images (left) and quantification (right) of invaded cells are shown. Data are presented as the mean ± standard deviation (SD) of three separate experiments, n = 3. ** p < 0.01, *** p < 0.001, and **** p < 0.0001 according to 2-tailed t test.

Article Snippet: Anti-β-actin antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). pDR-GFP (#17617), pCBASceI (#26477) and Rictor shRNA plasmids (#1853 and #1854) were obtained from Addgene (Watertown, MA, USA).

Techniques: Knockdown, Western Blot, Expressing, shRNA, Transwell Invasion Assay, Stable Transfection, Transfection, Control, Standard Deviation

Journal: Oncology Research

Article Title: mTORC2 promotes pancreatic cancer progression and parp inhibitor resistance

doi: 10.32604/or.2023.029309

Figure Lengend Snippet: Rictor knockdown enhances DNA damage induced by the PARP inhibitor olaparib in PDAC cells. (A) PANC-1 cells expressing control (Ctrl) or Rictor shRNA were treated with the indicated concentrations of olaparib, followed by clonogenic survival analysis. (B, C) PANC-1 cells expressing control (Ctrl) or Rictor shRNA were treated with or without 20 μM olaparib for 24 h, followed by western blot analysis of γH2AX (B) and comet DNA damage assay (C). Data are presented as the mean ± standard deviation (SD) of three separate experiments, n = 3. ** p < 0.01 according to 2-tailed t test.

Article Snippet: Anti-β-actin antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). pDR-GFP (#17617), pCBASceI (#26477) and Rictor shRNA plasmids (#1853 and #1854) were obtained from Addgene (Watertown, MA, USA).

Techniques: Knockdown, Expressing, Control, shRNA, Western Blot, Standard Deviation

Journal: Oncology Research

Article Title: mTORC2 promotes pancreatic cancer progression and parp inhibitor resistance

doi: 10.32604/or.2023.029309

Figure Lengend Snippet: Knockdown of Rictor suppresses HR-mediated repair. (A) Diagram of HR-mediated repair reporter, in which a GFP gene is interrupted by insertion of endonuclease I-SceI cut site; the functional GFP can be restored only by HR-mediated repair using the downstream iGFP sequence as a template. (B) HR-mediated repair capacity was measured in U2OS-DR-GFP cells transfected with control (Ctrl) or Rictor shRNA. (C) PANC-1 cells expressing control (Ctrl) or Rictor shRNA were treated with 20 μM olaparib, followed by immunofluorescence analysis of Rad51 foci. Data are presented as the mean ± standard deviation (SD) of three separate experiments, n = 3. ** p < 0.01 according to 2-tailed t test.

Article Snippet: Anti-β-actin antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). pDR-GFP (#17617), pCBASceI (#26477) and Rictor shRNA plasmids (#1853 and #1854) were obtained from Addgene (Watertown, MA, USA).

Techniques: Knockdown, Functional Assay, Sequencing, Transfection, Control, shRNA, Expressing, Immunofluorescence, Standard Deviation

Journal: Oncology Research

Article Title: mTORC2 promotes pancreatic cancer progression and parp inhibitor resistance

doi: 10.32604/or.2023.029309

Figure Lengend Snippet: Rictor interacts with BRCA1. (A) Coimmunoprecipitation (Co-IP) analysis of the Rictor/BRCA1 association in PANC-1 cells using an anti-Rictor antibody. (B) PANC-1 cells expressing control (Ctrl) or Rictor shRNA were treated with 20 μM olaparib, followed by analysis of BRCA1 foci. Data are presented as the mean ± standard deviation (SD) of three separate experiments, n = 3. *** p < 0.001 according to 2-tailed t test.

Article Snippet: Anti-β-actin antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). pDR-GFP (#17617), pCBASceI (#26477) and Rictor shRNA plasmids (#1853 and #1854) were obtained from Addgene (Watertown, MA, USA).

Techniques: Co-Immunoprecipitation Assay, Expressing, Control, shRNA, Standard Deviation