Notes: Kaiso functions as either a transcriptional repressor or activator in pathways related to cell cycle control, cancer progression, and apoptosis. Here, Kaiso is shown to be monoSUMOylated on lysine 42 under normal cellular conditions and functions as a repressor. DeSUMOylation of Kaiso leads to a functional switch from acting as a repressor to an activator. The Dual-Luciferase® Reporter Assay is used to monitor expression under the control of Kaiso. (5087)

Notes: The presence of post-transcriptional modifications (PTMs) in viral RNAs is investigated using mass spectrometry. A wide-range of PTMs on RNAs from Zika virus, Dengue virus, HIV-1, Poliovirus, and hepatitis C virus were identified. Additionally, PTMs on cellular RNAs were measured in response to viral infection and stress response. The Dual-Luciferase® Reporter Assay was used to monitor cellular stress. (5091)

Notes: The authors studied the ability of JAK1 V658F and A634D mutants to activate the Janus kinase (JAK)/STAT pathway when expressed alone or together with the other components of the interleukin-9 receptor complex. The BOX1 motif of wild-type IL-9Rα, the JAK interacting region, was mutated from PXP to SXS using the GeneEditor™ in vitro Site-Directed Mutagenesis System. To assess STAT transcriptional activity, HEK293 human embryonic kidney, COS-7 monkey kidney, U4C human fibrosarcoma and g2A cells were cotransfected with 250ng of the appropriate constructs, 500ng of firefly luciferase vectors and 50ng of pRL-TK Vector and empty plasmid for a total 1.5µg of DNA. After 24 hours, the cells were lysed in 150µl of Passive Lysis Buffer and reporter activity measured using the Dual-Luciferase^® Reporter Assay System. The ProFection^® Mammalian Transfection System—Calcium Phosphate was used to transfect 10⁶ HEK293 cells in a six-well plate with 3.75µg of plasmid for Western blot analysis and cotransfected 6 × 10⁶ HEK293 cells in a 100mm dish with 14µg plasmid for immunoprecipitation studies. (4025)

Notes: The authors used real-time quantitative PCR to characterize cytokine response after HIV infection of human lymphoid tissues. To synthesize first-strand cDNA, total RNA was reverse transcribed using the ImProm-II™ Reverse Transcription System: 200U of ImProm-II Reverse Transcriptase, 2µg of total RNA, 500ng of oligo(dT) primer, 500µM dNTPs 3mM MgCl₂ and 24U of RNase inhibitor in 1x ImProm-II™ reaction buffer. To obtain viral stocks for infection, 293T cells were transfected with the proviral plasmids pNL4-3 and pYU-2 using the ProFection^® Mammalian Transfection System—Calcium Phosphate. (3455)

Notes: The authors seek to determine the role of glutamate in excitotoxicity and neurofilament accumulation seen in some neurodegenerative diseases. Neurofilament light, middle, and heavy chains were expressed from rat cDNAs cloned into the pCI-neo Mammalian Expression vector in SW-13 cells. Primary rat cortical neurons were transfected with a neurofilament middle chain and green fluorescent fusion protein. SW13 cells and primary rat cortical neurons were transfected with the ProFection® Mammalian Transfection System–Calcium Phosphate. The CytoTox 96® Non-Radioactive Cytotoxicity Assay was used to monitor glutamate toxicity in these cell. To determine the role of the MAPK and JNK signaling pathways,  SW13- cells and primary neuronal cells were immunostained for dually phosphorylated MAPK and JNK using Promega's Anti-ACTIVE® MAPK pAb and Anti-ACTIVE® JNK pAb, respectively. Cells were fixed in 4% paraformaldehyde, permeabilized in 0.1% Triton® X-100 in PBS, blocked with  0.2% Tween® 20 in TBS, and incubated with primary antibodies diluted in blocking solution. Western blot analyses were performed on the primary cortical neurons to quantitate the level of dually phosphorylated MAPK protein  The blots were also probed with a pan MAPK antibody that detects total (active and inactive) MAPK. (2382)

Notes: The ProFection® Mammalian Transfection System - Calcium Phosphate was used to transfect HeLa and HeLa-Tat cell lines. The authors developed a vector-antiviral gene system for use in gene therapy in the treatment of HIV-1. pGEM®-luc was used as a negative control in luciferase assays of transfected cells. (1388)

Notes: The ProFection^® Mammalian Transfection System-Calcium Phosphate was used to transfect reporter plasmids into HuH-7, a human hepatoma cell line. The cells were plated at 1.2 X 10⁶ in a 100mm dish and transfected 48 hours later. Transfection efficiency was monitored with a co-transfection of the pSV-β-Galactosidase Control Vector with the CAT reporter vectors. (1130)

Notes: ProFection^® Mammalian Transfection System (Calcium Phosphate) was used to transfect primary cerebellar granule neurons from 8 day old Sprague Dawley rats. The complete details of the transfection is provided, including the efficiency of the transfection. Efficiencies, as determined by transfected control β-galactosidase, ranged from 3-7%. (0170)

Notes: Transfections of HepG2 cells, 2.2.15 (a Hepatitis B-infected cell line) and HeLa cells were accomplished with the ProFection^® Mammalian Transfection System-Calcium Phosphate. Transfection efficiencies were monitored with co-transfected pSV-β-Galactosidase Control Vector, but the efficiencies were not reported. (0843)

Notes: The Profection^® Mammalian Transfection System (CaPO₄) was used for transient transfection of the Drosophila Schneider line 2 cells. A lot of detail is provided. (1230)

Notes: The ProFection^® Mammalian Transfection System-CaPO₄ was used to transiently transfect 293T cells. (0521)

Notes: The inhibitor was used to protect RNA during RT-PCR. (2246)

Notes: The ProFection^® Mammalian Transfection System-CaPO₄ transfection system was used to generate stably transfected ML-RCC renal carcinoma cells. (0628)

Notes: The ProFection^® Mammalian Transfection System-Calcium Phosphate was used to transiently transfect HeLa-T4+ cells at 50% confluency. (0138)