Journal: Cell Death & Disease

Article Title: The DNA methylation-regulated miR-193a-3p dictates the multi-chemoresistance of bladder cancer via repression of SRSF2/PLAU/HIC2 expression

doi: 10.1038/cddis.2014.367

Figure Lengend Snippet: The effect of both miR-193a and its target genes on the BCa chemoresistance. ( a ) The experimental scheme. ( b ) The relative cell survival by the IC 50 -dosed indicated drugs in the miR-193a-3p (3PM) or miR-193a-5p (5PM) mimic versus the mock (NC) of 5637 cells in plot. ( c ) The relative level of both SRSF2 (-3p target) and TP73 (-5p target) normalized with the GAPDH level, in the same set of 5637 cells in panel ( b ). ( d ) The relative cell survival by the IC 50 -dosed indicated drugs in miR-193a-3p (3PA) or miR-193a-5p (5PA) antagomiR versus the mock (NC) in H-bc cells in plot. ( e ), The relative level of SRSF2 (-3p target) and TP73 (-5p target) proteins normalized with the GAPDH level, in the same set of H-bc cells in panel ( d ). ( f ) The relative cell survival of the siRNA or 3PM-transfected or NC-transfected 5637 cells under the IC 50 -dosed Pa and Ad, respectively. ( g ) The protein level of the downstream genes in the same set of cells in panel ( f ), by western blotting analysis. ( h ) The relative cell survival of 5637 cells transfected with the indicated transfection under the IC 50 -dosed Pa and Ad, respectively. ( i ) The protein level of the experiment in panel ( h ) (Pa-treated group), by western blotting analysis. ( j ) The summary of the effects on the cell survival of the IC 50 -dosed drug-trigger cell death by miR-193a-3p's target genes. ( k ) The relative cell survival under the treatment of the IC 50 -dosed Pa and Ad in H-bc cells transfected with the SRSF2-GFP, PLAU-GFP expression constructs, 3PA or NC, respectively. (* P <0.05; ** P <0.01)

Article Snippet: Briefly, the cells were transfected in triplet with each firefly luciferase reporter construct in combination with the Renilla luciferase construct using ribo FECT CP transfection reagent, and both luciferase activities in cell extracts at 24 h after transfection were measured by a Promega Dual-Luciferase Reporter assay (Promega) using a Promega GloMax 20/20 luminometer.

Techniques: Transfection, Western Blot, Expressing, Construct

Journal: SpringerPlus

Article Title: Proliferative effect and osteoinductive potential of extracellular matrix coated on cell culture plates

doi: 10.1186/2193-1801-2-303

Figure Lengend Snippet: Transfection with shBMP-2 affected ALP activity and Runx2 mRNA level of MC3T3-E1 cells re-seeded on osteoblast-ECM. A , shBMP-2 reduced BMP-2 protein level of MC3T3-E1 cells. B , shBMP-2 lowered ALP activity of MC3T3-E1 cells seeded on cell culture plates without ECM coating (no ECM) , but had little effect on MC3T3-E1 cells seeded on osteoblast-ECM coated plates (ECM). C , shBMP-2 reduced Runx2 mRNA level, but had little effect on the cells seeded on osteoblast-ECM. ** P <0.01, *P <0.05, between the indicated groups, n = 5 per group.

Article Snippet: MC3T3-E1 pre-osteoblastic cells (60%-70% confluence) were transfected with shBMP2 (shFGF2) or control shRNA (shCtl) using High-Fect Transfection Reagent (Beijing Cowin Bioscience Co., Ltd., Beijing, China), according to the manufacturer’s recommendations.

Techniques: Transfection, Activity Assay, Cell Culture

Journal: SpringerPlus

Article Title: Proliferative effect and osteoinductive potential of extracellular matrix coated on cell culture plates

doi: 10.1186/2193-1801-2-303

Figure Lengend Snippet: Transfection with shFGF-2 affected proliferation of MC3T3-E1 cells seeded on fibroblast-ECM. A , shFGF-2 reduced FGF-2 protein level of MC3T3-E1 cells. B , shFGF-2 lowered proliferative activity of MC3T3-E1 cells seeded on cell culture plates without ECM coating (no ECM), but had little effect on MC3T3-E1 cells seeded on fibroblast-ECM coated plates. ** P <0.01, *P <0.05, between the indicated groups, n = 5 per group.

Article Snippet: MC3T3-E1 pre-osteoblastic cells (60%-70% confluence) were transfected with shBMP2 (shFGF2) or control shRNA (shCtl) using High-Fect Transfection Reagent (Beijing Cowin Bioscience Co., Ltd., Beijing, China), according to the manufacturer’s recommendations.

Techniques: Transfection, Activity Assay, Cell Culture