Journal: FASEB BioAdvances

Article Title: Role of alternatively spliced, pro‐survival Protein Kinase C delta VIII (PKCδVIII) in ovarian cancer

doi: 10.1096/fba.2021-00090

Figure Lengend Snippet: Overexpression of PKCδVIII in T80 increases cell migration. (A) T80 cells grown to confluency with μ‐Dish inserts (Ibidi solutions, 81176) and were transfected with 2 μg pTracer‐PKCδVIII plasmid for 48 h. Inserts were removed to make consistent gaps and cells were imaged at 0, 6, and 24 h. The red lines are drawn on the image to aid in visualization of the edges of the gaps. The cells were imaged using Keyence BZx‐810 microscope and analysis of gap area between cells (μm 2 ) was quantified using FastTrack A1 software (Ibidi solutions™). (B) RNA was then isolated from the cells and PCR performed using PKCδVIII specific primers. The products were separated on a 1% agarose gel and stained using ethidium bromide. Graph shows relative quantification of PKCδVIII normalized to β‐actin with control set as reference. The experiments were independently repeated three times with similar results. Statistical analysis was performed using two‐way ANOVA; *** p < 0.001

Article Snippet: The ovarian cancer cells with and without the PKCδVIII siRNA were allowed to migrate for 18 h. After 18 h, the cells that migrated to the bottom well were fixed and stained using crystal violet and imaged on Keyence BZx‐810 microscope.

Techniques: Over Expression, Migration, Transfection, Plasmid Preparation, Microscopy, Software, Isolation, Agarose Gel Electrophoresis, Staining

Journal: FASEB BioAdvances

Article Title: Role of alternatively spliced, pro‐survival Protein Kinase C delta VIII (PKCδVIII) in ovarian cancer

doi: 10.1096/fba.2021-00090

Figure Lengend Snippet: PKCδVIII associates with Bcl2 and Bcl‐xL in ovarian cells. Two micrograms of pTracer‐PKCδVIII was transfected in T80 cells for 48 h. (A) Western blot analysis was performed on input lysates and probed using antibodies as indicated. Coimmunoprecipitation assay was performed on the cell lysates using antibodies against (B) PKCδVIII (IP: PKCδVIII, n = 4) (C) Bcl2 (IP: Bcl2, n = 3), (D) Bcl‐xL (IP: Bcl‐xL, n = 3), and (e) BAD (IP:BAD, n = 4). Western blot analysis was then performed on the immunoprecipitated samples (IP) using antibodies against PKCδVIII, Bcl2, Bcl‐xL, and Bad as indicated in the figure. Graphs show densitometric analysis of individual band normalized to immunoprecipitated antibody target. Statistical analysis was performed using t ‐test; ** p < 0.01, *** p < 0.001, and ns = not significant. The experiments were independently repeated three times with similar results. (F) Immunocytochemistry was performed on T80, SKOV3, and TOV112D ovarian cells by fixing and staining cells using antibodies against PKCδVIII, Bcl2, Bcl‐xL, Bad, and DAPI (for nucleus) and as indicated in the figure. Cells were imaged using a Keyence BZx‐810 microscope and Pearson's colocalization was calculated using Keyence Analyzer software. The experiments were independently repeated four times with similar results. Statistical analysis was performed using one‐way ANOVA; *** p < 0.001 and ns = not significant

Article Snippet: The ovarian cancer cells with and without the PKCδVIII siRNA were allowed to migrate for 18 h. After 18 h, the cells that migrated to the bottom well were fixed and stained using crystal violet and imaged on Keyence BZx‐810 microscope.

Techniques: Transfection, Western Blot, Co-Immunoprecipitation Assay, Immunoprecipitation, Immunocytochemistry, Staining, Microscopy, Software

Journal: FASEB BioAdvances

Article Title: Role of alternatively spliced, pro‐survival Protein Kinase C delta VIII (PKCδVIII) in ovarian cancer

doi: 10.1096/fba.2021-00090

Figure Lengend Snippet: Depletion of PKCδVIII in ovarian cancer cells decreases Bcl2 and cellular migration, viability, and proliferation. 25 nM of PKCδVIII specific siRNA was transfected in SKOV3 (SKOV3 PKCδVIIIsi ) and TOV112D (TOV112D PKCδVIIIsi ) or 25nM control siRNA (control) was transfected into the cells for 48 h. (A) RNA was isolated and Real‐time SYBR Green qPCR was performed for PKCδVIII, PKCδI, Bcl2, and β‐actin. Graphs show relative quantification of PKCδVIII, Bcl2, and Bcl‐xL normalized to β‐actin with control set as reference. The experiments were independently repeated four times with similar results. Statistical analysis was performed using t ‐test; ** p < 0.01, *** p < 0.001 and ns = not significant. (B) Whole cell lysates were harvested and Western blot was performed using antibodies as indicated. The experiments were independently repeated four times with similar results. (C) SKOV3 and TOV112D were grown to confluency with μ‐Dish inserts (Ibidi solutions, 81176), PKCδVIII was depleted by siRNA transfection for 48 h. Inserts were removed to make consistent gaps and cells were imaged at 0, 24, and 48 h. The red lines are drawn on the image to aid in visualization of the edges of the gaps. The experiments were independently repeated six times with similar results. Cells were imaged using Keyence BZx‐810 microscope and analysis of gap area between cells (μm 2 ) was quantified using FastTrack A1 software (Ibidi solutions™). Statistical analysis was performed using two‐way ANOVA; ** p < 0.01 and *** p < 0.001. (D) Cell migration assay using a transwell insert was performed as described in methods. After 18 h, cells in the bottom well were stained with crystal violet and imaged using Keyence BZx‐810 microscope. Cells were counted using the Keyence Analyzer software. The experiments were independently repeated three times with similar results. Statistical analysis was performed using one‐way ANOVA; *** p < 0.001. (E) Cell viability was determined using WST1 assay by measuring the formazon dye produced using a plate reader. The experiments were independently repeated four times with similar results. Statistical analysis was performed using t ‐test; *** p < 0.001. (F) Cell proliferation was determined using BrdU assay by incubating cells in BrdU for 2 h, then fixing and staining with anti‐BrdU antibody. Cells were then stained with peroxidase secondary antibody and measured by plate reader. The experiments were independently repeated four times with similar results. Statistical analysis was performed using t ‐test; *** p < 0.001

Article Snippet: The ovarian cancer cells with and without the PKCδVIII siRNA were allowed to migrate for 18 h. After 18 h, the cells that migrated to the bottom well were fixed and stained using crystal violet and imaged on Keyence BZx‐810 microscope.

Techniques: Migration, Transfection, Isolation, SYBR Green Assay, Western Blot, Microscopy, Software, Cell Migration Assay, Staining, Produced, BrdU Staining

Journal: Journal of NeuroEngineering and Rehabilitation

Article Title: Devices used for photobiomodulation of the brain—a comprehensive and systematic review

doi: 10.1186/s12984-024-01351-8

Figure Lengend Snippet: Categories and parameters of the reviewed devices

Article Snippet: Vielight Intranasal 810 (Vielight Inc., CA) [ , , ] , 3 , Intranasal LED , 1 LED diode , Ventromedial prefrontal, entorhinal cortex, hippocampus , 1 , 810 , 10 Hz 50% DC , 14.20 , 10.65 , 10.65 , 0.014 , AD; Dementia; Gulf War Illness.

Techniques:

Journal: Journal of NeuroEngineering and Rehabilitation

Article Title: Devices used for photobiomodulation of the brain—a comprehensive and systematic review

doi: 10.1186/s12984-024-01351-8

Figure Lengend Snippet: Comparison of device categories

Article Snippet: Vielight Intranasal 810 (Vielight Inc., CA) [ , , ] , 3 , Intranasal LED , 1 LED diode , Ventromedial prefrontal, entorhinal cortex, hippocampus , 1 , 810 , 10 Hz 50% DC , 14.20 , 10.65 , 10.65 , 0.014 , AD; Dementia; Gulf War Illness.

Techniques: Comparison