Journal: Molecular Medicine Reports

Article Title: Lentivirus-mediated RIG-I knockdown relieves cell proliferation inhibition, cell cycle arrest and apoptosis in ATRA-induced NB4 cells via the AKT-FOXO3A signaling pathway in vitro

doi: 10.3892/mmr.2017.6858

Figure Lengend Snippet: Expression of RIG-I in ATRA-induced NB4 cells is knocked down by LV-mediated shRIG-I. (A) Flow cytometric detection of the expression of GFP in LV-shCon and LV-shRIG-I cells at 72 h post-lentivirus infection. (B) Reverse transcription-quantitative polymerase chain reaction analysis for detection of the RNA expression of RIG-I in NB4, LV-shCon and LV-shRIG-I cells at 0 and 72 h post-ATRA induction (n=3). *P<0.01. (C) Western blot analysis of the protein expression of RIG-I in NB4, LV-shCon and LV-shRIG-I cells at 0 and 72 h post-ATRA induction. (D) Quantitative analysis of protein expression of RIG-I in NB4, LV-shCon and LV-shRIG-I cells at 0 and 72 h post-ATRA induction (n=3) *P<0.01. Data are presented as the mean ± standard deviation. RIG-I, retinoic acid inducible gene I; ATRA, all-trans retinoic acid; GFP, green fluorescent protein; LV, lentivirus; Con, control; siRNA, small interfering RNA; LV-shCon, NB4 cells infected with LV Con-siRNA; LV-shRIG-I, NB4 cells infected with LV RIG-I-siRNA.

Article Snippet: Subsequently, lentivirus containing green fluorescent protein (GFP) (Shanghai Genepharma Co., Ltd, Shanghai, China) and polybrene (final concentration of 8 μg/ml) were added to each well of NB4 cells, which were then centrifuged at 1,000 g for 90 min at room temperature.

Techniques: Expressing, Infection, Reverse Transcription, Real-time Polymerase Chain Reaction, RNA Expression, Western Blot, Standard Deviation, Control, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: Lentivirus-mediated RIG-I knockdown relieves cell proliferation inhibition, cell cycle arrest and apoptosis in ATRA-induced NB4 cells via the AKT-FOXO3A signaling pathway in vitro

doi: 10.3892/mmr.2017.6858

Figure Lengend Snippet: RIG-I knockdown alleviates ATRA-induced reduced proliferation and cell cycle arrest in NB4 cells. (A) MTT detection of NB4, LV-shCon and LV-shRIG-I cell proliferation without ATRA induction at different time points. (B) MTT detection of NB4, LV-shCon and LV-shRIG-I cell proliferation with ATRA induction at different time points (n=3) # P<0.05. (C) Flow cytometric detection of NB4, LV-shCon and LV-shRIG-I cells in the G1 phase at 0 and 72 h post-ATRA induction (n=3) # P<0.05. Data are presented as the mean ± standard deviation. RIG-I, retinoic acid inducible gene I; ATRA, all-trans retinoic acid; LV, lentivirus; Con, control; siRNA, small interfering RNA; LV-shCon, NB4 cells infected with lentivirus Con-siRNA; LV-shRIG-I, NB4 cells infected with lentivirus RIG-I-siRNA. NB4 + ATRA, ATRA-induced NB4 cells; LV-shCon + ATRA, ATRA-induced LV-shCon cells; LV-shRIG-I + ATRA, ATRA-induced LV-shRIG-I cells.

Article Snippet: Subsequently, lentivirus containing green fluorescent protein (GFP) (Shanghai Genepharma Co., Ltd, Shanghai, China) and polybrene (final concentration of 8 μg/ml) were added to each well of NB4 cells, which were then centrifuged at 1,000 g for 90 min at room temperature.

Techniques: Knockdown, Standard Deviation, Control, Small Interfering RNA, Infection

Journal: Molecular Medicine Reports

Article Title: Lentivirus-mediated RIG-I knockdown relieves cell proliferation inhibition, cell cycle arrest and apoptosis in ATRA-induced NB4 cells via the AKT-FOXO3A signaling pathway in vitro

doi: 10.3892/mmr.2017.6858

Figure Lengend Snippet: RIG-I knockdown inhibits ATRA-induced apoptosis in NB4 cells. (A) Flow cytometric detection of apoptosis in NB4, LV-shCon and LV-shRIG-I cells at 0 and 72 h post-ATRA induction. (B) Quantitative analysis of apoptosis in NB4, LV-shCon and LV-shRIG-I cells at 0 and 72 h post-ATRA induction (n=3) *P<0.01. Data are presented as the mean ± standard deviation. RIG-I, retinoic acid inducible gene I; ATRA, all-trans retinoic acid; LV, lentivirus; Con, control; siRNA, small interfering RNA; LV-shCon, NB4 cells infected with lentivirus Con-siRNA; LV-shRIG-I, NB4 cells infected with lentivirus RIG-I-siRNA.

Article Snippet: Subsequently, lentivirus containing green fluorescent protein (GFP) (Shanghai Genepharma Co., Ltd, Shanghai, China) and polybrene (final concentration of 8 μg/ml) were added to each well of NB4 cells, which were then centrifuged at 1,000 g for 90 min at room temperature.

Techniques: Knockdown, Standard Deviation, Control, Small Interfering RNA, Infection

Journal: Molecular Medicine Reports

Article Title: Lentivirus-mediated RIG-I knockdown relieves cell proliferation inhibition, cell cycle arrest and apoptosis in ATRA-induced NB4 cells via the AKT-FOXO3A signaling pathway in vitro

doi: 10.3892/mmr.2017.6858

Figure Lengend Snippet: RIG-I-knockdown reduces cell proliferation inhibition, cell cycle arrest and apoptosis in ATRA-induced NB4 cells via the AKT-FOXO3A signaling pathway. (A) Western blot analyses of expression levels of pAKT-Thr308, pFOXO3A-Thr32, p27 and TRAIL in LV-shCon and LV-shRIG-I cells at 0 and 72 h post-ATRA induction. (B) Quantitative analysis of protein levels of pAKT-Thr308 in LV-shCon and LV-shRIG-I cells at 0 and 72 h post-ATRA induction (n=3) *P<0.01. (C) Quantitative analysis of protein levels of pFOXO3A-Thr32 in LV-shCon and LV-shRIG-I cells at 0 and 72 h post-ATRA induction (n=3) *P<0.01. (D) Quantitative analysis of protein levels of p27 in LV-shCon and LV-shRIG-I cells at 0 and 72 h post-ATRA induction (n=3) *P<0.01. (E) Quantitative analysis of protein levels of TRAIL in LV-shCon and LV-shRIG-I cells at 0 and 72 h post-ATRA induction (n=3) *P<0.01. Data are presented as the mean ± standard deviation. RIG-I, retinoic acid inducible gene I; ATRA, all-trans retinoic acid; LV, lentivirus; Con, control; siRNA, small interfering RNA; LV-shCon, NB4 cells infected with lentivirus Con-siRNA; LV-shRIG-I, NB4 cells infected with lentivirus RIG-I-siRNA.

Article Snippet: Subsequently, lentivirus containing green fluorescent protein (GFP) (Shanghai Genepharma Co., Ltd, Shanghai, China) and polybrene (final concentration of 8 μg/ml) were added to each well of NB4 cells, which were then centrifuged at 1,000 g for 90 min at room temperature.

Techniques: Knockdown, Inhibition, Western Blot, Expressing, Standard Deviation, Control, Small Interfering RNA, Infection

Journal: Frontiers in Pharmacology

Article Title: Licochalcone A suppresses pancreatic ductal adenocarcinoma progression by targeting eEF2K-mediated pyroptosis

doi: 10.3389/fphar.2025.1595686

Figure Lengend Snippet: LHA induces pyroptosis through targeting eEF2K in PDAC cells. (A) Molecular docking of LHA on eEF2K protein. Predicted two-dimensional and three-dimensional crystal structures of LHA (PubChem CID: 5318998) and the eEF2K (PDB ID: 7SHQ) complex. (B) MIA PaCa-2 and BxPc3 cell lines were treated with LHA (0, 7.5, 10, and 12.5 μM) for 48 h. The expressions of eEF2K, p-eEF2, and eEF2 were examined using Western blotting. Data were expressed as mean ± S.D. of at least three independent experiments. *p < 0.05, **p < 0.01, and ***p < 0.001 vs. control. (C) MIA PaCa-2 and BxPc3 cell lines were transfected using lentivirus vectors with the overexpressed human eEF2K gene, and the images were captured using the fluorescent microscope after 72 h of transfection. (D) MIA PaCa-2 and BxPc3 with or without eEF2K overexpression. The expressions of eEF2K were examined using Western blotting. Data were expressed as mean ± S.D. of at least three independent experiments. **p < 0.01 vs. control. (E) The cell viability of MIA PaCa-2 and BxPc3 cell lines with or without overexpression of eEF2K was examined using MTT assay. Data were expressed as mean ± S.D. of at least three independent experiments. *p < 0.05, **p < 0.01, and ***p < 0.001 vs. control. # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. overexpression eEF2K with the same LHA concentration. (F) MIA PaCa-2 and BxPc3 with or without eEF2K overexpression with LHA (0 and 12.5 μM) for 48 h. The expressions of eEF2K, GSDMD-N, GSDME-N, and IL-1β were examined using Western blotting. Data were expressed as mean ± S.D. of at least three independent experiments. *p < 0.05, **p < 0.01, and ***p < 0.001 vs. control. ## p < 0.01 and ### p < 0.001 vs. overexpression eEF2K with the same LHA concentration.

Article Snippet: Lentivirus vectors carrying the overexpressed human eEF2K gene and green fluorescent protein (GFP) were designed and produced as “LV-EEF2K (100354-1).” The construct was bought from the Genchem Company (Shanghai, China).

Techniques: Western Blot, Control, Transfection, Microscopy, Over Expression, MTT Assay, Concentration Assay