Journal: Cancer Cell International

Article Title: RelB plays an oncogenic role and conveys chemo-resistance to DLD-1 colon cancer cells

doi: 10.1186/s12935-018-0677-x

Figure Lengend Snippet: Establishment of RelB-silencing DLD1 cells. a Western blotting analysis of the protein expression of NF-κB subunits in DLD-1, HT-29, and Caco-2 cells. The level of each protein was normalized against Actin. b Relative RelB mRNA expression levels were detected using qRT-PCR. β-Actin normalized gene expression, measured in triplicates was displayed. c Western blotting analysis for protein levels of RelB expression. Protein expression was normalized against Actin. d RelB-silencing affects the expression of other individual NF-κB subunits. Western blotting analysis of the protein expression in cytoplasmic and nuclear extracts was normalized against Actin and Lamin A/C, respectively. e Western blotting analysis of the protein expression of IKKβ, p-IKKβ, IKBα, and IKKα in siRelB and sictrl cells. Protein levels were normalized against Actin. f The DNA-binding activity of nuclear extracts was detected and quantified using a Trans AM NF-κB family transcription factor assay kit. g The mRNA expression levels of specific target genes downstream of NF-κB pathway were detected using qRT-PCR. β-Actin normalized gene expression, measured in triplicates was displayed. Data are shown as mean ± SD form three individual experiments. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Abs against NF-κB p65 (C-20, sc-372), RelB (C-19, sc-226), c-Rel (N, sc-70), NF-κB p105/50 (H-119, sc-7178), NF-κB p100/52 (K-27, sc-298), IKBα (H-4), IKKα (B-8),and Lamin A/C (H-110, sc-20681) were purchased from Santa Cruz Biotechnology, lnc.

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Gene Expression, Binding Assay, Activity Assay, Transcription Factor Assay

Journal: Bioengineering & Translational Medicine

Article Title: Neuron‐targeted 2‐deoxyglucose‐dendrimer‐rosiglitazone nanotherapy mitigates neuroinflammation and cognitive deficits in pediatric traumatic brain injury

doi: 10.1002/btm2.70053

Figure Lengend Snippet: FIGURE 8 mRNA expressions of pro- and anti-inflammatory markers and cell death markers from sham (n = 12, 6 M/6 F), traumatic brain injury (TBI) + vehicle (n = 12, 6 M/6 F), TBI + rosiglitazone (Rosi) (n = 12, 6 M/6 F), and TBI + 2-deoxyglucose (2DG-D)-Rosi (n = 12, 6 M/6 F) groups. Neurons were isolated from the injured brain regions (or the matching area from the sham mice) at 24-h post-treatment for gene expression evaluations. Data were presented as mean ± SEM. Treatment groups were presented as sham (gray circles), TBI + vehicle (blue circles), TBI + Rosi (magenta circles), and TBI + 2DG-D-Rosi (green circles). (a–d) The expression of pro-inflammatory makers, tumor necrosis factor-alpha (TNF-α) (a), interleukin-6 beta (IL-1β) (b), Toll-like receptor 4 (TLR4) (c), and NLR Family Pyrin Domain Containing 3 (NLRP3) (d). (e) The expression of anti-inflammatory maker IL-13. (f) The expression of cell death marker Fas. *p < 0.05; **p < 0.01; ***p < 0.001. (g) The co- localization of neurodegenerative markers marker Fluoro-Jade C (FJC) (green) and neuronal marker (NeuN, red) was evaluated in sham (n = 8, 4 M/4 F), TBI + vehicle (n = 8, 4 M/4 F), TBI + Rosi (n = 8, 4 M/4 F), and TBI + 2DG-D-Rosi (n = 8, 4 M/4 F) groups at 24-h post-injury. Images (40, 5 images/animal) were randomly acquired from the cortex area (mainly primary motor cortex and primary somatosensory cortex) in the injured brain regions (approximately between bregma +1 mm and bregma 0.5 mm) using the Nikon Eclipse TS2R fluorescent microscope (Nikon, NY, USA). The representative images from male treatment groups (upper panels) and female treatment groups (lower panels). 40,6-Diamidino-2-phenylindole (DAPI) (blue) was used for nucleus stains. Scale bars: 100 μm. (h) The quantification of FJC+NeuN+ cells among treatment groups. Data were presented as mean ± SEM. Treatment groups were presented as sham (gray circles), TBI + vehicle (blue circles), TBI + Rosi (magenta circles), and TBI + 2DG-D-Rosi (green circles). *p < 0.05; **p < 0.01; ****p < 0.0001.

Article Snippet: Brain sections were washed 3 times in PBS for 5 min each, incubated in 0.001% Fluoro-Jade C (FJC) (Cat# TR-160-FJC, Biosensis, CA, USA) for 10 min, washed six times in PBS for 15 min each, and incubated with rabbit anti-NeuN (1:250; Cat# ab177487; Abcam, MA, USA).

Techniques: Isolation, Gene Expression, Expressing, Marker, Microscopy

Journal: International journal of molecular sciences

Article Title: Surgical Site-Released Tissue Is Potent to Generate Bone onto TCP and PCL-TCP Scaffolds In Vitro.

doi: 10.3390/ijms242115877

Figure Lengend Snippet: Figure 6. (a–c) ELISA boxplots of the osteogenic markers osteocalcin (a), osteopontin (b), and collagen I (c). Boxplots compare the concentration of ocn, opn, and col I in the control group with the PCL-TCP and β-TCP group over the cultivation time of 6 weeks. Groups are further subdivided into the condition’s agarose and glue, to examine if the chosen adhesive, used for the attachment of the scaffolds onto the plastic wells, makes a difference in protein expression. The boxblots indicating the median within the 25–75% percentile. The dots represent single outliners during measurements.

Article Snippet: Enzyme-linked immunosorbent assays (ELISAs) (human Duoset ELISA kit, R&D Systems, Bio Techne, Minneapolis, MN, USA) were then performed using the supernatants at a dilution of 1:10 for osteocalcin (1:50; pure for collagen and pure for osteopontin) according to the manufacturer’s protocol and measured at 450 nm using a Multiscan Ascent reader (Thermo Scientific).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Control, Adhesive, Expressing