nih 3t3 whole cell lysate  (Santa Cruz Biotechnology)

Journal: International Journal of Dentistry

Article Title: Acinar Cell Proliferation Promoted by BMP2 in Injured Mouse Parotid Gland: BMP2 Promotes Cell Proliferation in Parotid Gland

doi: 10.1155/2023/1765317

Figure Lengend Snippet: Mouse parotid acinar cells in primary culture. (a) Cell proliferative capacity of BMP2-added (100 ng/mL) and nonadded (control) groups after 48 hr of culture. The proliferative capacity of BMP2-added cultured cells was significantly increased compared to that in the control group. ∗∗ Indicates significance at P < 0.01, ∗ P < 0.05. Data were shown as mean ± SD. Three independent experiments were performed. (b) Protein expression of E-cadherin (an epithelial marker) and vimentin (a mesenchymal marker) in cultured cells of each group at 48 hr after the addition of BMP2. NIH3T3 (3T3) was used as a positive control for mesenchymal markers. RS: rat serum.

Article Snippet: NIH3T3 whole cell lysate (Novus Biologicals, Centennial, CO, USA) was used as a positive control for mesenchymal markers.

Techniques: Control, Cell Culture, Expressing, Marker, Positive Control

Journal: BMC Cancer

Article Title: Identification of Makorin 1 as a novel SEREX antigen of esophageal squamous cell carcinoma

doi: 10.1186/1471-2407-9-232

Figure Lengend Snippet: Isolation of stable MKRN1 transfectants . (a) Protein expression levels of MKRN1, His-tagged proteins, ubiquitinated proteins and β-actin in MKRN1 transfectants, Fmkrn-24 and 21, and the parental ras-NIH3T3 cells were examined by Western blotting using the anti-MKRN1 peptide antibody, the anti-His antibody (lower panel), the anti-ubiquitin antibody or the anti-β-actin antibody. (b) Expression of MKRN1 mRNA in MKRN1 transfectant Fmkrn-24 and ras-NIH3T3 cells was examined by RT-PCR. MKRN1 sequence between nucleotide positions 29 and 536 was amplified. The position of PCR products is indicated by an arrow. (c) Phase morphology of ras-NIH3T3 and Fmkrn-24 cells. Original magnification, ×100.

Article Snippet: Extracts of MKRN1-transfected ras-NIH3T3 cells were immunoprecipitated with anti-ubiquitin antibody (Santa Cruz) followed by SDS-polyacrylamide gel electrophoresis as described previously [ ].

Techniques: Isolation, Expressing, Western Blot, Transfection, Reverse Transcription Polymerase Chain Reaction, Sequencing, Amplification

Journal: BMC Neuroscience

Article Title: Activation of Erk and JNK MAPK pathways by acute swim stress in rat brain regions

doi: 10.1186/1471-2202-5-36

Figure Lengend Snippet: Swim stress did not induce phosphorylation of MKK3/6. Note the absence of a band for P-MKK3/6 at ~40 kD, either in control or swim stressed hippocampal lysates, whereas an induced band is visible in the positive control cell extracts. Also note the unknown ~37 and 48 kD proteins induced by swim stress. Other brain regions examined exhibited a similar pattern. 3T3, lysates of NIH/3T3 cells treated with (+) or without (-) UV radiation (40 mJ, 45 minutes recovery time); C6, lysates of C6 glioma cells treated with (+) or without (-) anisomycin (25 μg/ml, 30 min); MW, molecular weight markers.

Article Snippet: Control cell extracts for P-MKK3/6 were obtained from CST: +/- UV-treated NIH/3T3 cell lysates (9233) and +/- anisomycin-treated C6 glioma cell lysates (9213).

Techniques: Positive Control, Molecular Weight

Journal: Molecular Cancer

Article Title: Activation of Akt pathway by transcription-independent mechanisms of retinoic acid promotes survival and invasion in lung cancer cells

doi: 10.1186/1476-4598-12-44

Figure Lengend Snippet: Inhibition of the PI3k/Akt pathway promotes apoptosis by activation of caspase-3. ( A ) Left, A549 cells were serum-starved and treated or non-treated (control) with ATRA for 48 h, during the first 12 h after treatment with ATRA, the cells were irradiated with 150 J/m 2 of UV-C light for 30 min. Subsequently, DNA fragmentation was detected by TUNEL according to the manufacturer’s instructions. The apoptotic cells are stained brown. Bar, 20 μm. Right, percentages of TUNEL-positive cells were quantified by counting 200 cells from four random microscopic fields (means ± SEM, * P < 0.05 compared with non-treated cells (control) assessed by t test analysis). ( B ) A549 cells were treated for 48 h with 5 μM of ATRA alone or combined with 5 μM of 15e. Subsequently, DNA fragmentation was detected by TUNEL. Control cells were non-treated. Percentages of TUNEL-positive cells were quantified by counting 200 cells from four random microscopic fields. Means ± SEM, * P < 0.05; ** P < 0.001 compared with non-treated cells (control) (analysis of variance and Newman-Keuls test). ( C ) A549 cells were serum-starved and treated or non-treated (control) with 5 μM of ATRA alone or combined with 5 μM of 15e for 48 h. The cells were fixed, stained with anti-cleaved caspase-3 followed by donkey anti-goat FITC as described in Materials and Methods and analyzed by fluorescence microscopy. Bar, 20 μm.

Article Snippet: In some experiments, cells were incubated with anti-RARα (MCA4135Z; Serotec) and anti-Akt (P-2482; Sigma-Aldrich) or anti-cleaved caspase-3 (sc-22171; Santa Cruz) followed by incubation with anti-mouse Alexa Fluor 532, anti-mouse Alexa fluor 647 or anti-goat FITC (sc-2024; Santa Cruz), respectively.

Techniques: Inhibition, Activation Assay, Irradiation, TUNEL Assay, Staining, Fluorescence, Microscopy