Journal: Scientific Reports

Article Title: TDP-43 self-interaction is modulated by redox-active compounds Auranofin, Chelerythrine and Riluzole

doi: 10.1038/s41598-018-20565-0

Figure Lengend Snippet: TDP-43 self-interaction. NanoBit ® luciferase complementation assay for protein interactions has been used to measure interaction of different N-terminal large (LgBit, pFN33), small (SmBit, pFN35), C-terminal large (LgBit, pFC34) and small (SmBit, pFC36) fusion proteins of TDP-43, positive control vectors SmBiT-PRKACA and LgBiT-PRKAR2A (coding the catalytic and regulatory subunits of PKA) and vector SmBiT-Halotag, which contains haloalkane dehalogenase – smBit fusion protein (Halitag) as a negative control, were obtained from Promega (Madison, WI, USA) and have been used in parallel 24 h after transfection of N2a cells and seeding cells into 384-well plates, Mean ± SD, Mann-Whitney-U-Test **p < 0.01.

Article Snippet: NanoBit ® luciferase complementation assay for protein interactions, N-terminal large (LgBiT, pFN33) and small (SmBiT, pFN35), C-terminal LgBiT (pFC34) and SmBiT (pFC36) vectors, constitutive NanoLuc luciferase pNL1.1.TK[Nluc/TK] vector ( = Nluc vector), positive control vectors SmBiT-PRKACA and LgBiT-PRKAR2A and negative control vector SmBiT-Halotag were obtained from Promega (Madison, WI, USA).

Techniques: Luciferase, Positive Control, Plasmid Preparation, Negative Control, Transfection, MANN-WHITNEY

Journal: Cell & Bioscience

Article Title: Tumor protein D52 is upregulated in oral squamous carcinoma cells under hypoxia in a hypoxia-inducible-factor-independent manner and is involved in cell death resistance

doi: 10.1186/s13578-021-00634-0

Figure Lengend Snippet: Effects of knockdown and overexpression of TPD52 on growth and apoptosis of SAS cells. a , b siRNA for TPD52 or control siRNA was transfected into SAS cells, and incubated under normoxic conditions for 24 h. Then, the cells were exposed to hypoxia in the presence or absence of 10 μM PX-478. After 0, 1, 2, and 3 d cells were subjected to MTT and caspase 3/7 assays ( a ), and after 0 and 3 d, total cellular proteins were subjected to western blotting analysis to determine expression of TPD52, HIF-1α, p62, Akt, p-Akt, and β-actin ( b ). c , d HaloTag-TPD52 or control HaloTag vector was transfected into SAS cells, and incubated under normoxic conditions for 24 h. Then, the cells were exposed to hypoxia in the presence or absence of 10 μM PX-478. After 0, 1, 2, and 3 d, cells were subjected to MTT and caspase 3/7 assays (c), and after 0 and 3 d, total cellular proteins were subjected to western blotting analysis to detect TPD52 (overexpressed and endogenous proteins indicated with arrows with the molecular weight), HIF-1α, p62, Akt, p-Akt, HaloTag, and β-actin ( d ). For MTT and caspase 3/7 assays, the value at time 0 is designated as “1,” and relative values are shown. The values of 3 d were subjected to ANOVA. *, p < 0.05. Experiments were repeated 3 times

Article Snippet: The expression vectors of HaloTag-TPD52 (pFN21AE3730) and HaloTag control vector (G659) were purchased from Promega.

Techniques: Over Expression, Transfection, Incubation, Western Blot, Expressing, Plasmid Preparation, Molecular Weight