pkr 3  (MedChemExpress)

Journal: Journal of Virology

Article Title: Exacerbated Apoptosis of Cells Infected with Infectious Bursal Disease Virus upon Exposure to Interferon Alpha

doi: 10.1128/JVI.00364-18

Figure Lengend Snippet: List of primers used for RT-qPCR

Article Snippet: Antibodies used in this study were rabbit polyclonal sera specific for IBDV VP3 ( 85 ), ISG-56 (catalog number sc-82946; Santa Cruz Biotechnology), Mx1/2/3 (catalog number sc-50509; Santa Cruz Biotechnology), PKR (catalog number sc-6282; Santa Cruz Biotechnology), p-PKR (catalog number T446; Abcam), total eIF2α (catalog number sc-11386; Santa Cruz Biotechnology), p-eIF2α (catalog number 44728G; Invitrogen), PARP (catalog number 9542; Cell Signaling), NF-κB–p65 (catalog number ab16502; Abcam), histone H2B (catalog number ab52484; Abcam), α-tubulin (catalog number 2125; Cell Signaling), ubiquitin (catalog number sc-8017; Santa Cruz Biotechnology), and β-actin (catalog number sc-47778; Santa Cruz Biotechnology).

Techniques:

Journal: Molecular Vision

Article Title: PKR promotes choroidal neovascularization via upregulating the PI3K/Akt signaling pathway in VEGF expression

doi:

Figure Lengend Snippet: Effect of PKR siRNA and PI3K inhibitor LY294002 on hypoxia-induced PKR, p-PI3K, p-Akt, and VEGF expression in RF/6A cells. A : DsRNA-activated protein kinase (PKR), phosphophosphatidylinositol 3-kinase (p-PI3K), phosphoprotein kinase B (p-Akt), and vascular endothelial growth factor (VEGF) expression was detected with western blotting in the RF/6A cells following PKR siRNA transfection after 48 h. C : The histogram shows the densitometric analysis of the average levels for PKR, p-PI3K, p-Akt, and VEGF to GAPDH. Cells transfected with scramble siRNA were used as the negative control. B : The western blot shows PKR, p-PI3K, p-Akt, and VEGF expression in RF/6A cells treated with the PI3K inhibitor LY294002 for 30 min. D : The histogram shows the densitometric analysis of the average levels of PKR, p-PI3K, p-Akt, and VEGF to GAPDH. Hypoxic cells were used as the negative control. *p<0.05, statistically significantly different compared to the respective controls. Values represent means ± SD.

Article Snippet: Transfection of PKR siRNA (5′-GGA TTC GGG TTA CTT GTA A-3′, RiboBio, Guangzhou, China) was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA) following the manufacturer’s protocol.

Techniques: Expressing, Western Blot, Transfection, Negative Control

Journal: Molecular Vision

Article Title: PKR promotes choroidal neovascularization via upregulating the PI3K/Akt signaling pathway in VEGF expression

doi:

Figure Lengend Snippet: PKR promotes migration of RF/6A cells in a coculture system under hypoxic conditions. A : Crystal violet staining detected the migrated cells of each group: normal, hypoxic, scramble siRNA, and dsRNA-activated protein kinase (PKR) siRNA. Representative photographs of migrated RF/6A cells (200× magnification). B : The average number of migrated RF/6A cells per field. *p<0.01, PKR siRNA group versus the hypoxic group. Values represent means ± SD.

Article Snippet: Transfection of PKR siRNA (5′-GGA TTC GGG TTA CTT GTA A-3′, RiboBio, Guangzhou, China) was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA) following the manufacturer’s protocol.

Techniques: Migration, Staining

Journal: Molecular Vision

Article Title: PKR promotes choroidal neovascularization via upregulating the PI3K/Akt signaling pathway in VEGF expression

doi:

Figure Lengend Snippet: PKR promotes tube formation of RF/6A cells in a coculture system under hypoxic conditions. A : Microscopic images showing tube formation in each group: normal, hypoxic, scramble siRNA, and dsRNA-activated protein kinase (PKR) siRNA. Representative photographs of tube formation of RF/6A cells (200× magnification). B : Decreased tube formation was observed when the cocultured RF/6A cells were transfected with PKR siRNA under hypoxia. *p<0.01, PKR siRNA group versus the hypoxia group. Values represent means ± SD.

Article Snippet: Transfection of PKR siRNA (5′-GGA TTC GGG TTA CTT GTA A-3′, RiboBio, Guangzhou, China) was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA) following the manufacturer’s protocol.

Techniques: Transfection

Journal: BMC Genomics

Article Title: HIV-1 gp120 influences the expression of microRNAs in human monocyte-derived dendritic cells via STAT3 activation

doi: 10.1186/s12864-015-1673-3

Figure Lengend Snippet: HIV gp120 up-regulates PIAS3 and STAT1 protein expression in MDDCs. MDDCs were stimulated with gp120 (5 μg/ml) for 18 h. Cell lysates were resolved by 8-12 % SDS-PAGE, transferred to a nitrocellulose membrane, and subjected to immunoblot analysis with antibodies specific for: p38, ERK1/2, JNK, p65, p50, IkBα, IKKβ, JAK2, STAT-1, −2, −3, SOCS-1, −3, PIAS3, Bcl2, BCLxL, Akt, and PKR. Data from one representative experiment are shown. a Graphs show the level of protein as determined by densitometry (ImageJ software) and calculated relatively to untreated control ( C ), where each sample was normalized to total actin. The average fold from two-four independent experiments was represented, along with the SD. b Immunoblot analysis with antibodies specific for PIAS3 and STAT1. Actin expression is shown as gel loading control. Values below the lanes show band intensities of the respective bands, normalized to actin expression. Data from one representative experiment out of four analyzed are shown

Article Snippet: MDDCs were stimulated with gp120 (5 μg/ml) for 18 h. Cell were lysed in RIPA buffer (150 mM NaCl, 50 mM Tris-Cl (pH 7.5), 1 % Nonidet P-40, 0.5 % sodium deoxycholate, and 0.1 % SDS) containing a cocktail of protease and phosphatase inhibitors and protein extracts were resolved by 8-12 % SDS-PAGE, transferred to a nitrocellulose membrane, and subjected to immunoblot analysis with antibodies specific for STAT-1, STAT −3, SOCS1, NF-kB p50, IkBα, IKKβ, p38, ERK1/2, JNK, Akt and PIAS3 (Cell Signaling Technology; catalog numbers: 9172, 9139, 3950, 3035, 9242, 2684, 9212, 9102, 9252, 9272, 9042), and SOCS3, JAK2, STAT-2, NF-kB p65, Bcl2, BCLxL and PKR (Santa Cruz; catalog numbers: sc51699, sc476, sc81334, sc509, sc634, sc634, sc707), and actin (BD Transduction Laboratories; catalog numbers: 612656).

Techniques: Expressing, SDS Page, Membrane, Western Blot, Software, Control