cationic lipid transfections  (Thermo Fisher)

Journal: Oncology Letters

Article Title: Knockdown of spindle pole body component 25 homolog inhibits cell proliferation and cycle progression in prostate cancer

doi: 10.3892/ol.2018.8003

Figure Lengend Snippet: SPC25 knockdown inhibits the proliferation of PC-3 cells. (A) Expression of SPC25 mRNA following transfection with the indicated shRNAs in PC-3 cells. (B) An MTT assay demonstrated that knockdown of SPC25 inhibited cell proliferation in PC-3 cells. (C) The Celigo ® system revealed that knockdown of SPC25 inhibited cell proliferation in PC-3 cells. (D) Cell number of shCtrl or shSPC25 group in each day was calculated using the Celigo ® system. P

Article Snippet: Opti-modified Eagle's medium (Opti-MEM) which was ideal for use during cationic lipid transfections especially Lipofectamine 2000 transfection reagents was purchased from Thermo Fisher Scientific, Inc (cat. no. 31985062).

Techniques: Expressing, Transfection, MTT Assay

Journal: The Prostate

Article Title: Overexpression of MCM10 promotes cell proliferation and predicts poor prognosis in prostate cancer. Overexpression of MCM10 promotes cell proliferation and predicts poor prognosis in prostate cancer

doi: 10.1002/pros.23703

Figure Lengend Snippet: MCM10 knockdown inhibits its expression levels in DU145 and PC‐3 cells. A‐C, Real‐time reverse transcription PCR analysis showed expression of MCM10 mRNA was reduced after transfection with the indicated shRNAs in DU145 (A) PC‐3 (B) and LNCaP (C) cells. D‐F, Western blot analysis showed protein expression of MCM10 was reduced after transfection with the indicated shRNAs in DU145 (D), PC‐3 (E), and LNCaP (F) cells. Data are represented as the mean ± SD (* P

Article Snippet: Opti‐modified Eagle's medium (Opti‐MEM) which was ideal for use during cationic lipid transfections especially Lipofectamine™ transfection reagents was purchased from Thermo Fisher Scientific, Inc. (Catalog number: 31985062).

Techniques: Expressing, Polymerase Chain Reaction, Transfection, Western Blot

Journal: The Prostate

Article Title: Overexpression of MCM10 promotes cell proliferation and predicts poor prognosis in prostate cancer. Overexpression of MCM10 promotes cell proliferation and predicts poor prognosis in prostate cancer

doi: 10.1002/pros.23703

Figure Lengend Snippet: MCM10 knockdown inhibits its expression levels in DU145 and PC‐3 cells. A‐C, Real‐time reverse transcription PCR analysis showed expression of MCM10 mRNA was reduced after transfection with the indicated shRNAs in DU145 (A) PC‐3 (B) and LNCaP (C) cells. D‐F, Western blot analysis showed protein expression of MCM10 was reduced after transfection with the indicated shRNAs in DU145 (D), PC‐3 (E), and LNCaP (F) cells. Data are represented as the mean ± SD (* P

Article Snippet: Opti‐modified Eagle's medium (Opti‐MEM) which was ideal for use during cationic lipid transfections especially Lipofectamine™ transfection reagents was purchased from Thermo Fisher Scientific, Inc. (Catalog number: 31985062).

Techniques: Expressing, Polymerase Chain Reaction, Transfection, Western Blot

Journal: PLoS Pathogens

Article Title: Vesicular trafficking permits evasion of cGAS/STING surveillance during initial human papillomavirus infection

doi: 10.1371/journal.ppat.1009028

Figure Lengend Snippet: HPV16 evades cGAS/STING responses in HaCaTs during initial infection. cGAS/STING responses to pGL3 and HPV PsV in HaCaTs. (A, B) Cells were either transfected for 90 min with 250 ng pGL3 or infected for the duration of the experiment with 950 ng L1 (equivalent to 250 ng DNA) of HPV16 virions. Cells were harvested at various times post-treatment and cGAS/STING responses assessed via western blotting. (A) IRF3 was phosphorylated, most prominently at 90 min and 4 hr post pGL3 transfection, while IRF3 was not phosphorylated at any time post HPV16 infection, representative blot of n = 4 biological replicates. (B) Densitometric quantification of western blots, n = 4 biological replicates. Statistics calculated by two-way ANOVA ( P interaction = 0.0138) followed by Sidak’s multiple comparison test (*** P

Article Snippet: The calculated packaging ratio of 11.16 was therefore used to normalize luciferase data in experiments comparing pGL3 delivery via cationic lipid transfection to wildtype HPV infection (Figs and ).

Techniques: Infection, Transfection, Western Blot

Journal: PLoS Pathogens

Article Title: Vesicular trafficking permits evasion of cGAS/STING surveillance during initial human papillomavirus infection

doi: 10.1371/journal.ppat.1009028

Figure Lengend Snippet: HPV-related DNA-responsive cellular genes are not induced by virion infection. Normalized RNA-seq count data were plotted for a subset of 15 DNA-responsive cellular genes selected based on prior literature indicating a relationship with HPV. Error bars represent 95% confidence intervals ( n = 2 for pGL3 transfection, red, and n = 3 for HPV virion infection, black).

Article Snippet: The calculated packaging ratio of 11.16 was therefore used to normalize luciferase data in experiments comparing pGL3 delivery via cationic lipid transfection to wildtype HPV infection (Figs and ).

Techniques: Infection, RNA Sequencing Assay, Transfection

Journal: PLoS Pathogens

Article Title: Vesicular trafficking permits evasion of cGAS/STING surveillance during initial human papillomavirus infection

doi: 10.1371/journal.ppat.1009028

Figure Lengend Snippet: HPV16 virion infection does not activate cellular responses to dsDNA. RNA-seq was used to transcriptionally profile cellular responses to pGL3 DNA introduced via liposome transfection or HPV virion infection. (A) Relative to H 2 O mock-transfection, there were 142 significantly ( P -adj

Article Snippet: The calculated packaging ratio of 11.16 was therefore used to normalize luciferase data in experiments comparing pGL3 delivery via cationic lipid transfection to wildtype HPV infection (Figs and ).

Techniques: Infection, RNA Sequencing Assay, Transfection

Journal: PLoS Pathogens

Article Title: Vesicular trafficking permits evasion of cGAS/STING surveillance during initial human papillomavirus infection

doi: 10.1371/journal.ppat.1009028

Figure Lengend Snippet: HPV16 evades cGAS/STING responses in HFKs during initial infection. cGAS/STING responses to pGL3 and wildtype HPV in primary HFKs. (A) HFKs were either transfected for 90 min with 500 ng pGL3 or infected for the duration of the experiment with 1900 ng L1 (equivalent to 500 ng DNA) of HPV virions. Cells were harvested at various times post-treatment and cGAS/STING responses assessed via western blotting. (A) IRF3 was phosphorylated 4 hr post pGL3 transfection, while IRF3 was not phosphorylated at any time post HPV infection with HPV PsV. representative blot of n = 3 biological replicates. (B) Cells were either transfected for 90 min with 250 ng pGL3 or infected for the duration of the experiment with an equivalent amount of HPV virions. Luciferase activity was measured 24 hr post-treatment, normalized to GAPDH, and corrected for pGL3 encapsidation as described in Materials and Methods . * P

Article Snippet: The calculated packaging ratio of 11.16 was therefore used to normalize luciferase data in experiments comparing pGL3 delivery via cationic lipid transfection to wildtype HPV infection (Figs and ).

Techniques: Infection, Transfection, Western Blot, Luciferase, Activity Assay