The 24ST1NLESG cell line of HIV-1 latency was kindly provided by Dr. Joseph P. Dougherty (Robert Wood Johnson Medical School). Analysis of HIV-1 latency reversal in the 24ST1NLESG cells was conducted as described previously [14 (link)]. Briefly, in the 24ST1NLESG cell line, the integrated HIV-1 genome encodes a secretable alkaline phosphatase (SEAP) gene in env, which serves as an indicator of late gene expression, and used a measure of latency reversal. PF-3758309 was purchased from Selleckchem (Houston, TX, USA). siRNA molecules were obtained from DharmaconTM (Horizon Discovery Ltd., Co., USA). All other reagents were of the highest quality available and were used without further purification.
Pf 3758309
Manufactured by Selleck Chemicals
Sourced in United States
The PF-3758309 is a piece of laboratory equipment designed for general research and analytical purposes. It is a versatile instrument that can be used to perform a variety of tasks in a laboratory setting. The core function of the PF-3758309 is to facilitate the analysis and investigation of samples and materials.
Automatically generated - may contain errors
Lab products found in correlation
Frax597, by Selleck Chemicals (2 mentions)
Tamoxifen, by Merck Group (2 mentions)
Trametinib, by Selleck Chemicals (2 mentions)
4 oht, by Merck Group (2 mentions)
Imatinib, by Selleck Chemicals (2 mentions)
Murine anti pd l1 mab anti pd l1 migg1e3 invivofit, by InvivoGen (2 mentions)
Nintedanib, by Selleck Chemicals (2 mentions)
Azd4547, by Selleck Chemicals (2 mentions)
Kpt9274, by Selleck Chemicals (2 mentions)
Eclipse te300 microscope, by Nikon (1 mentions)
Endothelial cell medium (ecm), by ScienCell (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Bxpc 3, by American Type Culture Collection (1 mentions)
Mia paca 2, by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
12 protocols using pf 3758309
1
HIV-1 Latency Reversal Assay
2
Exosome-mediated Endothelial Tube Formation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Endothelial cell tube formation assays were performed as previously described [42 (link)]. Briefly, 2F-2B and HUVEC (7 × 104 cells/well) were re-suspended in DMEM + 5% FBS, and seeded onto growth factor-reduced BD matrigel (1 mg/ml) (96-well) for 1 h. Cells were then supplemented for 2 h with MDCK, MDCKYBX1 or 21D1 Exos (30 μg), or the vehicle control (DMEM). For inhibitor based tube formation assays, 2F-2B and HUVEC cells (7 × 104 cells/well) were pre-treated with Rac1 inhibitors: 1 μM FRAX597 (Selleck Chem) and 1 μM PF-3758309 (Selleck Chem) for 1 h. Cells were isolated (480 x g. 5 min) and re-suspended in DMEM + 5% FBS, and seeded onto growth factor-reduced BD matrigel (1 mg/ml) (96-well) for 1 h. Cells were then supplemented with MDCK, MDCKYBX1 or 21D1 Exos (30 μg), or the vehicle control (DMEM). Tube-like structures were imaged using Nikon Eclipse TE300 microscope.
3
PAK Inhibitor Endogenous Inhibition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The PAK inhibitor, PF-3758309 (Selleckchem), was used for endogenous PAK inhibition at 10 μM for 90 min. PhosTag molecule was produced by MRC PPU Reagents and Services, University of Dundee. All solvents and chemicals used for LC-MS/MS analysis were of MS grade (Sigma).
4
Long-Term Clonogenic Proliferation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary mPDAC cell cultures were isolated from autochthonous PDAC and cultured as described previously59 (link)
. All cells were cultivated for less than 30 passages, authenticated by genotyping and tested for mycoplasma contamination by PCR. Conventional human PDAC cell lines and primary patient derived low passaged PDAC cell cultures were established and cultured as previously reported58 (link)
.
For long-term clonogenic cell proliferation assays, cells were seeded into 24-well plates (density of 1-2×103 cells/well, depending on growth rate. The following day, plates were treated with different concentrations of drugs as indicated. Every 7 days, media and drug were refreshed. Cells were fixed and stained with 0.2% crystal violet in an ethanol/water solution 7 to 13 days after the start of treatment, according to the confluence reached by the untreated control. Crystal violet was solubilized with 10% acetic acid and absorbance was quantified at 595 nm. The resulting values were used to calculate the Bliss synergy score with the online software Synergy Finder (v1.0)60 (link)
. All assays were performed independently at least three times. Trametinib, nintedanib, AZD-4547, imatinib and PF-3758309 were obtained from Selleckchem, 4-OHT from Sigma, murine anti PD-L1 mAb (Anti PD-L1-mIgG1e3 InvivoFit™) was purchased from InvivoGen, and tamoxifen for in vivo treatment from Sigma.
. All cells were cultivated for less than 30 passages, authenticated by genotyping and tested for mycoplasma contamination by PCR. Conventional human PDAC cell lines and primary patient derived low passaged PDAC cell cultures were established and cultured as previously reported58 (link)
.
For long-term clonogenic cell proliferation assays, cells were seeded into 24-well plates (density of 1-2×103 cells/well, depending on growth rate. The following day, plates were treated with different concentrations of drugs as indicated. Every 7 days, media and drug were refreshed. Cells were fixed and stained with 0.2% crystal violet in an ethanol/water solution 7 to 13 days after the start of treatment, according to the confluence reached by the untreated control. Crystal violet was solubilized with 10% acetic acid and absorbance was quantified at 595 nm. The resulting values were used to calculate the Bliss synergy score with the online software Synergy Finder (v1.0)60 (link)
. All assays were performed independently at least three times. Trametinib, nintedanib, AZD-4547, imatinib and PF-3758309 were obtained from Selleckchem, 4-OHT from Sigma, murine anti PD-L1 mAb (Anti PD-L1-mIgG1e3 InvivoFit™) was purchased from InvivoGen, and tamoxifen for in vivo treatment from Sigma.
5
Cultivation and Treatment of Human Brain Microvascular Endothelial Cells
6
Culturing Human Neuroblastoma Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human neuroblastoma cell lines were purchased from JENNIO Biological Technology (Guangzhou, China) within 5 years. All cells were maintained as monolayer cultures in RPMI-1640, Dulbecco's modified Eagles medium (DMEM) or DMEM/F12 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS) (Atlanta Biologicals, Lawrenceville, GA, USA), penicillin (100 U/ml) and streptomycin (100 µg/ml) (Sigma, St. Louis, MO, USA) in a humidified atmosphere of 5% CO2 at 37°C. All cells were tested routinely for Mycoplasma. The small-molecular inhibitor PF-3758309 was purchased from Selleck Chemicals (Houston, TX, USA).
7
Pancreatic Cancer Cell Line Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SW-1990, PSN-1, BxPC3 and PANC-1 cell lines were purchased from ATCC. MIA PaCa-2 and KP4 were purchased from Sigma-Aldrich and Accegen, respectively. MIA PaCa-2, PANC-1, SW-1990 were cultured in DMEM (Sigma-Aldrich) whereas PSN-1, BxPC3 and KP4 were cultured in RPMI1640 (Sigma-Aldrich). Both the Medias were supplemented with 10% Fetal Bovine Serum (Gibco) and 5% Penicillin-Streptomycin (Sigma-Aldrich). Cell lines were maintained at 37 °C with 5% CO2 in a cell culture incubator. Gemcitabine Hydrochloride was procured from Sigma Aldrich (catalogue number: G6423). PAK4 inhibitors (KPT-9274, catalogue number: S8444 and PF-3758309, catalogue number: S7094) were procured from Selleckchem.
8
Cytoskeletal Dynamics and Cellular Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anti–VE-cadherin (F-8) was from Santa Cruz Biotechnology. Anti-CD31 (89C2) was from Cell Signaling Technologies. 4′,6-Diamidino-2-phenylindole (DAPI), rhodamine phalloidin (1 mg/ml), Alexa Fluor 647 phalloidin (1 mg/ml), and Alexa Fluor–conjugated secondary antibodies were from Life Technologies. For antibody concentrations, see the “Immunofluorescence” and “Western blot and small GTPase activity assays” sections below. Rapamycin, alpelisib (BYL719), Y-27362 2HCl, trametinib (GSK1120212), batimastat (BB-94), EHT1864 2HCl, and PF-3758309 were from Selleckchem.
9
Long-Term Clonogenic Proliferation Assay
10
Evaluation of Small Molecule Inhibitors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The drugs employed in this study were as follows: dabrafenib (ApexBio, B1407‐50); XL888 (ApexBio, A4388‐25); fludarabine (Selleckchem, S1491); CHIR‐99021 HCl (CT99021) (Selleckchem, S2924); palbociclib (ApexBio, A8316); LDC000067 (ApexBio, B4754‐10); Ro 3306 (ApexBio, A8885‐10); roscovitine (ApexBio, A1723‐10); K03861 (Selleckchem, S8100); CHIR‐99021 (Selleckchem, S2924); dinaciclib (Selleckchem, S2768); FRAX597 (Selleckchem, S7271); PF‐3758309 (Selleckchem, S7094); AUY922 (Selleckchem, S1069); BIIB021 (Selleckchem, S1175); novobiocin (Selleckchem, S2492); 17‐DMAG (Selleckchem, S1142). All the drugs were dissolved in DMSO (Sigma D2650).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!