Journal: Open Life Sciences

Article Title: BuyangHuanwu Decoction attenuates cerebral vasospasm caused by subarachnoid hemorrhage in rats via PI3K/AKT/eNOS axis

doi: 10.1515/biol-2022-0071

Figure Lengend Snippet: Analysis of PI3K, AKT, and p-AKT expression levels by western blot and quantitative real-time PCR. (a) The blots showed that the SAH group had lower levels of PI3K, AKT, and p-AKT than the control group, and the graphs (b) show the ratio of expression compared with actin. BHD increased the expression of PI3K, AKT, and p-AKT as compared with the SAH group. (c) Quantitative real-time PCR analysis of PI3K, AKT, and p-AKT. Quantitative real-time PCR analysis of data was adjusted for GAPDH mRNA content ( n = 3) and examined in three separate quadruplicate experiments; data were shown as mean ± standard deviation ( n = 3); * P < 0.05, compared with the control group and # P < 0.05, compared with the SAH group, were used to indicate significance.

Article Snippet: Rabbit polyclonal anti-rat PI3K (1:1000; Wuhan Servicebio Biotechnology Co., Ltd.), Akt, and p-AKT antibodies were used and anti-actin (monoclonal antibody, 1:1,000; Servicebio) served as a control.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Angelicin inhibits liver cancer growth in vitro and in vivo

doi: 10.3892/mmr.2017.7219

Figure Lengend Snippet: Treatment with angelicin alters the protein expression levels of PI3K, p-Akt and total Akt in HepG2 and Huh-7 cells in vitro . (A) Representative blots demonstrating PI3K, p-Akt and total Akt protein expression levels in HepG2 and Huh-7 cells, following treatment with various concentrations of angelicin. (B) PI3K blots were semi-quantified using densitometry analysis of the protein bands and normalized to GAPDH. (C) p-Akt/Akt blots were semi-quantified using densitometry analysis of the protein bands. (D) Effects of LY294002 on angelicin-induced apoptosis. Cells were treated with the PI3K inhibitor LY294002 (3 mM) for 1 h prior to treatment with angelicin. The percentage of apoptotic cells following treatment with angelicin in the presence or absence of LY294002 was assessed using Annexin V-fluorescein isothiocyanate/propidium iodide staining and flow cytometry. Data are expressed as the mean ± standard deviation of 3 independent experiments. *P<0.05, **P<0.01 vs. the control group (0 µM angelicin); # P<0.05 vs. the DMSO group. PI3K, phosphatidylinositol-4,5-bisphosphate 3-kinase; p-, phosphorylated; DMSO, dimethyl sulfoxide; Akt, RAC-α serine/threonine-protein kinase.

Article Snippet: Polyclonal antibodies against PI3K (AF3242), Akt (AF6261), phosphorylated (p)-Akt (AF0016), apoptosis regulator BAX (Bax; AF0083) and Bcl-2 (AF6139) were purchased from Affinity Biologicals, Inc. (Ancaster, ON, Canada); antibodies against caspase-9 (9508T), caspase-3 (9665S), cytochrome c (4272S), Ki-67 (12075) and p-vascular endothelial growth factor receptor (VEGFR) 2 (9698) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).

Techniques: Expressing, In Vitro, Staining, Flow Cytometry, Standard Deviation, Control

Journal: Immunity, Inflammation and Disease

Article Title: Long noncoding RNA X‐inactive specific transcript (lncRNA XIST) inhibits hepatic insulin resistance by competitively binding microRNA‐182‐5p

doi: 10.1002/iid3.969

Figure Lengend Snippet: Effects of lncRNA XIST on IGF‐1R, 2‐DG6P, G6Pase, and PEPCK expressions and PI3K/Akt pathway in IR cells. (A) LncRNA XIST expression was detected by qRT‐PCR, and GAPDH was served as the internal reference. (B) The concentration of 2‐DG6P in LX‐2 cells was detected by 2‐DG uptake measurement kit. (C) The mRNA expression levels of G6Pase and PEPCK were determined by qRT‐PCR, and GAPDH was served as the internal reference. (D–E) Western blot was used to detect the G6Pase and PEPCK protein expressions, and GAPDH was served as the internal reference. (F and G) The protein expression of IGF‐1R was also assessed by western blot, and GAPDH was served as the internal reference. (H–J) Western blot was performed again to measure the protein expressions of PI3K, p‐PI3K, Akt, and p‐Akt, and GAPDH was served as the internal reference. All experiments were repeated three times to average. The data were presented as the mean ± standard deviation (SD) of three independent experiments; *** p < .001 versus Blank; ^ p < .05; ^^ p < .01; ^^^ p < .001 versus EVC. 2‐DG6P, 2‐deoxy‐d‐glucose‐6‐phosphate; EVC, empty vector control; G6Pase, glucose‐6‐phosphatase; GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase; IGF‐1R, insulin‐like growth factor‐1 receptor; IR, insulin resistance; PEPCK, phosphoenolpyruvate carboxykinase; PI3K, phosphatidylinositol 3‐kinase; qRT‐PCR, quantitative reverse transcription polymerase chain reaction; shXIST, XIST short hairpin RNA; XIST, X‐inactive specific transcript.

Article Snippet: The primary antibodies used were those against G6Pase (1:1000; Rabbit; ab93857; abcam, 40 kDa), PEPCK (1:1000; Rabbit; No. ABIN2855891; antibodies‐online, 71 kDa), p‐PI3K (1:1000; Rabbit; #029801; Alamo Laboratories Inc, 85 kDa), PI3K (1:1000; Rabbit; #4249; Cell Signaling Technology (CST), 110 kDa), p‐Akt (1:2000; Rabbit; #4060; CST, 60 kDa), Akt (1:1000; Rabbit; #4685; CST, 60 kDa), insulin‐like growth factor‐1 receptor (IGF‐1R, 1:1000; Rabbit; ab182408; abcam, 156 kDa) and GAPDH (1:10000; Rabbit; ab181602; abcam, 36 kDa).

Techniques: Expressing, Quantitative RT-PCR, Concentration Assay, Western Blot, Standard Deviation, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, shRNA

Journal: Immunity, Inflammation and Disease

Article Title: Long noncoding RNA X‐inactive specific transcript (lncRNA XIST) inhibits hepatic insulin resistance by competitively binding microRNA‐182‐5p

doi: 10.1002/iid3.969

Figure Lengend Snippet: Effects of miR‐182‐5p on IGF‐1R, 2‐DG6P, G6Pase and PEPCK expressions and PI3K/Akt pathway in IR cells. (A) MiR‐182‐5p expression was detected by qRT‐PCR, with U6 as the internal reference. (B) The miR‐182‐5p expression level in EVC + MC, XIST + MC, EVC + M, and XIST + M groups were detected by qRT‐PCR, with U6 serving as the internal reference. (C) The 2‐DG6P content was detected by 2‐DG uptake measurement kit. (D) After transfection, the mRNA expressions of G6Pase and PEPCK were determined by qRT‐PCR, with GAPDH serving as the internal reference. (E–H) The protein expressions of G6Pase, PEPCK, and IGF‐1R were measured by western blot, with GAPDH serving as the internal reference. (I–K) The PI3K/Akt pathway‐related proteins were also detected by western blot, with GAPDH serving as the internal reference. All experiments were repeated three times to average. The data were presented as the mean ± standard deviation (SD) of three independent experiments; + p < .05 versus Blank; * p < .05; *** p < .001 versus EVC + MC; ^^ p < .01; ^^^ p < .001 versus XIST + MC; ### p < .001 versus EVC + M. 2‐DG6P, 2‐deoxy‐d‐glucose‐6‐phosphate; EVC, empty vector control; GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase; G6Pase, glucose‐6‐phosphatase; IGF‐1R, insulin‐like growth factor‐1 receptor; IR, insulin resistance; M, miR‐182‐5p mimic; MC, mimic control; PEPCK, phosphoenolpyruvate carboxykinase; PI3K, phosphatidylinositol 3‐kinase; qRT‐PCR, quantitative reverse transcription polymerase chain reaction; XIST, X‐inactive specific transcript.

Article Snippet: The primary antibodies used were those against G6Pase (1:1000; Rabbit; ab93857; abcam, 40 kDa), PEPCK (1:1000; Rabbit; No. ABIN2855891; antibodies‐online, 71 kDa), p‐PI3K (1:1000; Rabbit; #029801; Alamo Laboratories Inc, 85 kDa), PI3K (1:1000; Rabbit; #4249; Cell Signaling Technology (CST), 110 kDa), p‐Akt (1:2000; Rabbit; #4060; CST, 60 kDa), Akt (1:1000; Rabbit; #4685; CST, 60 kDa), insulin‐like growth factor‐1 receptor (IGF‐1R, 1:1000; Rabbit; ab182408; abcam, 156 kDa) and GAPDH (1:10000; Rabbit; ab181602; abcam, 36 kDa).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Standard Deviation, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: The Effect of Tou Nong San on Transplanted Tumor Growth in Nude Mice

doi: 10.1155/2015/518454

Figure Lengend Snippet: Expression of p-PI3K protein in tumor tissues from control and treated mice (400×). TNSE significantly decreased the expression of p-PI3K ( P < 0.001). (a) NS group; (b) 1 g/kg TNSE group; (c) 2 g/kg TNSE group; (d) 4 g/kg TNSE group.

Article Snippet: RNase A and antibodies against p-PI3K (Tyr-368), p-AKT (Ser-473), p-mTOR (Ser-2448), p-p70s6k1 (Ser-424), cleaved Caspase-3, cleaved Caspase-9, and β -actin were purchased from Nanjing Keygen Biotec Co., Ltd (Nanjing, China).

Techniques: Expressing

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: The Effect of Tou Nong San on Transplanted Tumor Growth in Nude Mice

doi: 10.1155/2015/518454

Figure Lengend Snippet: The positive area of p-PI3K, p-AKT, p-mTOR, and p-p70 s6k1 and integrated option density (IOD) were determined. A: NS group; B: 1 g/kg TNSE group; C: 2 g/kg TNSE group; D: 4 g/kg TNSE group. Values given are the means ± SD for 8 tumor specimens in each group.

Article Snippet: RNase A and antibodies against p-PI3K (Tyr-368), p-AKT (Ser-473), p-mTOR (Ser-2448), p-p70s6k1 (Ser-424), cleaved Caspase-3, cleaved Caspase-9, and β -actin were purchased from Nanjing Keygen Biotec Co., Ltd (Nanjing, China).

Techniques:

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: The Effect of Tou Nong San on Transplanted Tumor Growth in Nude Mice

doi: 10.1155/2015/518454

Figure Lengend Snippet: Expression of p-PI3K, p-AKT, p-m-TOR, and p-p70 s6k1 proteins in xenograft. The effects of the treatment with TNSE on protein expression were measured in multiple samples ( n = 8) of xenografts by Western blot. Densitometric analysis was evaluated by Syngene G:BOX Chemi XR5 (English) and results were expressed as relative units. A: NS group; B: 1 g/kg TNSE group; C: 2 g/kg TNSE group; D: 4 g/kg TNSE group. Values given are the means ± SD for 8 tumor specimens in each group. * P < 0.05, ** P < 0.01, when compared to control group.

Article Snippet: RNase A and antibodies against p-PI3K (Tyr-368), p-AKT (Ser-473), p-mTOR (Ser-2448), p-p70s6k1 (Ser-424), cleaved Caspase-3, cleaved Caspase-9, and β -actin were purchased from Nanjing Keygen Biotec Co., Ltd (Nanjing, China).

Techniques: Expressing, Western Blot