Journal: BMC Cancer

Article Title: Relevance of cyclin D1b expression and CCND1 polymorphism in the pathogenesis of multiple myeloma and mantle cell lymphoma

doi: 10.1186/1471-2407-6-238

Figure Lengend Snippet: CCND1 polymorphism, monoallelic transcription and cyclin D1 mRNA level

Article Snippet: The DCS-6 monoclonal anti-cyclin D1 antibody (Ab, Pharmingen BD Biosciences n°556070) is directed against the entire cyclin D1 molecule and recognizes both isoforms; the sc-718 anti-cyclin D1 Ab (Santa Cruz Biotechnologies) is directed against the C-terminal part of the protein and is specific for cyclin D1a, the R3 Ab is directed against the C-terminal part of cyclin D1b and is specific for this isoform [ ].

Techniques:

Journal: BMC Cancer

Article Title: Relevance of cyclin D1b expression and CCND1 polymorphism in the pathogenesis of multiple myeloma and mantle cell lymphoma

doi: 10.1186/1471-2407-6-238

Figure Lengend Snippet: Cyclin D1a and cyclin D1b protein expression . A. Whole cell extracts were obtained from CD19+ cells of MCL patients, separated on SDS-PAGE and sequentially immunoblotted with anti-cyclin D1a (sc-718) or anti-cyclin D1b (R3) Abs. For loading controls, blots were also revealed with anti-β-tubulin Ab (sc-9104, not shown). B. Proteins were obtained from MCL cell lines and analyzed as described in A except that we used DCS-6 anti-cyclin D1 Ab which recognizes both isoforms.

Article Snippet: The DCS-6 monoclonal anti-cyclin D1 antibody (Ab, Pharmingen BD Biosciences n°556070) is directed against the entire cyclin D1 molecule and recognizes both isoforms; the sc-718 anti-cyclin D1 Ab (Santa Cruz Biotechnologies) is directed against the C-terminal part of the protein and is specific for cyclin D1a, the R3 Ab is directed against the C-terminal part of cyclin D1b and is specific for this isoform [ ].

Techniques: Expressing, SDS Page

Journal: Aging (Albany NY)

Article Title: Exogenous biological renal support ameliorates renal pathology after ischemia reperfusion injury in elderly mice

doi: 10.18632/aging.101899

Figure Lengend Snippet: Exogenous biological renal support increased renal cell proliferation in old IRI mice. ( A ) Representative images of renal EdU-positive cells in independent groups (600× magnification; red, EDU; green, LTL; blue, DAPI). ( B ) The percentages of EdU-positive cells in the kidneys of the old mice at 72 hours after IRI. The mice in the O: IRI group displayed more EdU-positive cells than in O: sham group. The percentage of EdU-positive cells was higher in the O-O: IRI group and the Y-O: IRI group than in the O: IRI group. No significant difference was found between the O-O: IRI group and the Y-O: IRI group. ( C ) Representative images of renal PCNA-positive tubular cells in independent groups (400× magnification). ( D ) The percentages of PCNA-positive tubular cells in the kidneys of the old mice at 72 hours after IRI. The mice in the O: IRI group had more PCNA-positive tubular cells than the O: sham group. The percentages of PCNA-positive tubular cells were higher in the O-O: IRI group and the Y-O: IRI group than in the O: IRI group. No significant difference was found between the O-O: IRI group and the Y-O: IRI group. ( E ) The levels of cyclin D1 and cyclin E1 in kidney extracts of the old IRI mice as measured by western blotting. Gels were performed under the same experimental conditions. ( F, G ) Quantitative analyses of the band densities of cyclin D1 and cyclin E1 protein expression. Data are presented as means ± SDs. ▲P < 0.05, ▲▲P < 0.01 vs. O: sham; *P < 0.05, ** P < 0.01 vs. O: IRI. SD, standard deviation.

Article Snippet: We transferred proteins to nitrocellulose (NC) membranes and probed the membranes with primary antibodies against the following proteins overnight at 4 °C, including beta-actin (Beijing Biosynthesis Biotechnology Co. 0061R; 1:2000), Pax2 (Proteintech, 21385; 1:800), vimentin (Abcam ab8069; 1:1,000), Kim1(R&D Systems, AF1817; 1:500), ERK1/2 (CST, #9102; 1:800), p-ERK1/2 (CST, #9101; 1:800), cyclin D1 (Proteintech 60186; 1:1,000), and cyclin E1 (Proteintech 11554; 1:1,000).

Techniques: Western Blot, Expressing, Standard Deviation

Journal: Cell Proliferation

Article Title: CCND1, NOP14 and DNMT3B are involved in miR‐502‐5p–mediated inhibition of cell migration and proliferation in bladder cancer

doi: 10.1111/cpr.12751

Figure Lengend Snippet: CCND1, DNMT3B and NOP14 were identified as direct downstream targets of miR‐502‐5p. A, Bioinformatic prediction analysis. MiRWalk and TargetScan online databases were used to predict potential downstream targets of miR‐502‐5p. B, RT‐qPCR assay. Ten candidate targets were selected, and the mRNA levels of these genes were measured. Only CCND1, DNMT3B and NOP14 were significantly downregulated in both cell lines. C, KEGG pathways analysis. KOBAS tool was used to analyse the pathways involved (representative pathways are presented). D, Dual‐luciferase reporter assay. The luciferase activity was significantly reduced after miR‐502‐5p mimics were transfected into the wild‐type group. However, no marked changes in the luciferase activity were observed in the mutated type group. E, Western blot assay. Protein levels of several candidate targets were measured after transfection with miR‐502‐5p mimics. F, Schematic diagram of the miR‐502‐5p–targeting region of CCND1, DNMT3B and NOP14 with seed matching. Error bars represent the SE obtained from three independent experiments; * P < .05

Article Snippet: After blocking with bovine serum albumin (Sango Biotech, Shanghai, China), the slides were incubated with anti‐Ki‐67, anti‐CCND1, anti‐NOP14 (Proteintech) and anti‐DNMT3B (Cell Signaling Technology) overnight at 4°C.

Techniques: Quantitative RT-PCR, Luciferase, Reporter Assay, Activity Assay, Transfection, Western Blot