Journal: The Journal of Neuroscience

Article Title: Nociceptor Translational Profiling Reveals the Ragulator-Rag GTPase Complex as a Critical Generator of Neuropathic Pain

doi: 10.1523/JNEUROSCI.2661-18.2018

Figure Lengend Snippet: Antibodies and chemicals used in the study

Article Snippet: Anti-p-ERK1/2 produced in rabbit , Cell Signaling Technology , 4366.

Techniques: Produced, cDNA Synthesis, SYBR Green Assay, Western Blot, RNA Sequencing, Software

Journal: PLoS ONE

Article Title: Green Tea Polyphenols Induce p53-Dependent and p53-Independent Apoptosis in Prostate Cancer Cells through Two Distinct Mechanisms

doi: 10.1371/journal.pone.0052572

Figure Lengend Snippet: Isogenic cells with lenti-virus vector background were generated by permanent knockdown of p53 in LNCaP cells [LNCaPshp53] and transfected with control vector, LNCaPshV cells. [ A ] Cell treated with 50 and 100 ng/ml camptothecin for 24 h were harvested, stained with PI and analyzed by flow cytometry to measure sub-G1 and G1 population. [ B ] Cells were treated with 50, 100 and 200 ng/ml of camptothecin for 24 h and stained with methylene blue. The intensity of methylene blue taken up by live cells was measured spectrophotometerically after eluting the dye in 0.1N HCl and compared with untreated cells. [ C ] Knockdown of p53 upregulated Akt/BAD signaling in prostate cancer cells. LNCaPshV and LNCaPshp53 cells were lysed and Western blotting was performed for p53, Akt, p-Akt (Ser473), BAD, p-BAD (Ser136) proteins. Actin was used as internal loading control. [ D ] Relative intensities of p-Akt and p-Bad protein in LNCaPshV and LNCaPshp53RNA cells where bands were normalized to actin and expressed in relative values compared to the native protein. The details are described in the materials and methods section.

Article Snippet: Antibodies for anti-caspase-9 (#9502), anti-caspase-3 (#9662), cleaved PARP (#9544), anti-c-IAP (#3130), anti-X-IAP (#2045), anti-SAPK/JNK (#9258), anti-p-JNK Thr183/Tyr185 (#9251), anti-p-Akt Ser473 (#4051), anti-BAD (#9292), anti-p-BAD Ser136 (#9295) and anti-VDAC (#4866) were purchased from Cell Signaling Technologies (Danvers, MA).

Techniques: Virus, Plasmid Preparation, Generated, Knockdown, Transfection, Control, Staining, Flow Cytometry, Western Blot

Journal: PLoS ONE

Article Title: Green Tea Polyphenols Induce p53-Dependent and p53-Independent Apoptosis in Prostate Cancer Cells through Two Distinct Mechanisms

doi: 10.1371/journal.pone.0052572

Figure Lengend Snippet: [ A ] Cells were treated with 20–80 µg/ml concentration of GTP for 24 h and Western blotting was performed for p53, Akt, p-Akt (Ser473), BAD, p-BAD (Ser136), p21/waf1, PUMA and Bax proteins. [ B ] Cells were treated with 40 µg/ml concentration of GTP for 3, 6, 9, 12, 24 and 48 h and Western blotting was performed for procaspase-9, cleaved caspase-9 and cleaved caspase-3 proteins. A typical actin blot demonstrates internal loading control. Cytochrome c release from mitochondria to cytosol was determined by Western blotting in GTP treated cells. A typical actin blot demonstrates internal loading control for cytosol whereas VDAC as internal loading control for mitochondria. [ D ] Cells were treated with 20 µM concentration of PI3K-Akt inhibitor LY294002 for 8 h and with 40 µg/ml GTP for 16 h alone, or LY294002 for 8 h followed by GTP treatment in combination followed by Western blotting for p-Akt, Akt, p-BAD and BAD proteins. The expressions of native proteins were considered as loading controls. [ D ] Cell death measurement was performed by photometric enzyme immunoassay Cell Death Detection ELISA kit. The bars represent mean±SD of at least two independent experiments each performed in duplicate, ** p <0.001 represents significant differences as compared to control group. [ E ] Light microscopic images of LNCaPshV and LNCaPshp53 cells treated with LY294002 and GTP alone or in combination. The details are described in the materials and methods section.

Article Snippet: Antibodies for anti-caspase-9 (#9502), anti-caspase-3 (#9662), cleaved PARP (#9544), anti-c-IAP (#3130), anti-X-IAP (#2045), anti-SAPK/JNK (#9258), anti-p-JNK Thr183/Tyr185 (#9251), anti-p-Akt Ser473 (#4051), anti-BAD (#9292), anti-p-BAD Ser136 (#9295) and anti-VDAC (#4866) were purchased from Cell Signaling Technologies (Danvers, MA).

Techniques: Concentration Assay, Western Blot, Control, Enzyme-linked Immunosorbent Assay