rod1 over expression (OriGene)
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rod1 over expression
Rod1 Over Expression, supplied by OriGene, used in various techniques. Bioz Stars score: 96/100, based on 1039 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rod1+over+expression/pCMV6-XL5+Mammalian+Expression+Vector/pm23024754-64-0-11
Average 96 stars, based on 1039 article reviews
Rod1 Over Expression, supplied by OriGene, used in various techniques. Bioz Stars score: 96/100, based on 1039 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rod1+over+expression/pCMV6-XL5+Mammalian+Expression+Vector/pm23024754-64-0-11
Average 96 stars, based on 1039 article reviews
rod1 over expression - by Bioz Stars,
2026-09
96/100 stars
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1) Product Images from "ROD1 is a seedless target gene of hypoxia-induced miR-210."
Article Title: ROD1 is a seedless target gene of hypoxia-induced miR-210.
Journal: PloS one
doi: 10.1371/journal.pone.0044651
Figure Legend Snippet: Figure 1. Increased or decreased miR-210 levels are associated to higher or lower ROD1 transcript levels in the RISC, respectively. HEK-293 were co-transfected with expression vectors for either miR-210 or a scramble sequence, and c-myc-Ago2 (A, miR-210 column and B). Alternatively, HEK-293 (46103/cm2) were co-transfected with anti-miR-210 or anti-scramble LNA-oligonucleotides and c-myc- Ago2 (A, anti-210 column and C). Then, c-myc antibody was used to immuno-precipitate c-myc-Ago2-containing complexes. A) miRNA levels in RISCs derived from transfected cells were assayed by qPCR. As expected, miR-210 was enriched or deprived in RISCs derived from cells in which miR-210 levels were up- or down-modulated, respectively, whereas other miRNAs did not show any significant modulation. Average values are expressed using a log2 scale. Green and red colors indicate down- or up-regulation, respectively (n = 3; p,0.01). B) ROD1 mRNA levels in RISCs derived from miR-210-enriched cells were assayed by qPCR. Background controls were represented by c-myc-immuno- precipitates derived from cells transfected with miR-210, but not mAgo2 (n = 7; *p,0.001). C) ROD1 mRNA levels in RISCs derived from
Techniques Used: Transfection, Expressing, Sequencing, Derivative Assay
Figure Legend Snippet: Figure 2. miR-210 and hypoxia decrease ROD1 expression. HEK-293 were transfected with plasmids encoding either miR-210 (miR-210) or a scramble sequence (scramble). Then, 24 and 48 hrs later, cells were collected and ROD1 protein and mRNA were assayed (A–C). Alternatively, HEK- 293 were infected with lentiviruses expressing anti-scramble or anti-miR-210 sponges and, 24 hrs later, cells were exposed to hypoxia for further 72 hrs. Then cells were collected and ROD1 protein and mRNA were assayed (D-F). A) A representative western blot is shown; TUB: a-tubulin. B) The bar graph represents average ROD1 protein expression levels, measured by densitometric analysis and normalized for a-tubulin levels (n = 4; *p,0.001). C) ROD1 mRNA levels were measured by qPCR (n = 3; *p,0.001). D) A representative western blot is shown; TUB: a-tubulin. E) Expression levels of ROD1 protein were evaluated by densitometric analysis and normalized for a-tubulin protein levels (n-normoxia = 9; n-hypoxia = 5; *p,0.001). F) ROD1 mRNA levels were measured by qPCR (n = 3; *p,0.05; #p,0.005). doi:10.1371/journal.pone.0044651.g002
Techniques Used: Expressing, Transfection, Sequencing, Infection, Western Blot
Figure Legend Snippet: Figure 3. miR-210 targets ROD1 directly. A) miR-210 (green) and ROD1 (red) complementarity. ROD1 transcript variant 6 (NM_001244898) is the longest ROD1 isoform and it was used for base numeration. B) RNA-duplexes prediction between miR-210 (green) and ROD1, EFNA3 and ISCU (red). Their calculated thermodynamic stabilities are indicated (mfe = minimum free energy). The mRNA sequences displayed are the following (gene name, RefSeq, bases, location): ROD1- isoform 6, NM_001244898, 1216–1236, cds; EFNA3, NM_004952,798– 804, 39UTR; ISCU, NM_014301, 111–118, 39UTR. C) HEK-293 were transfected with firefly luciferase constructs that contain either an intact (pLUC-ROD1-wt) or a mutated miR-210 binding site (pLUC-ROD1-del). pLUC plasmids were co-transfected with a plasmid encoding renilla luciferase along with a plasmid encoding either miR-210 (miR-210) or a scramble sequence (scramble). Firefly luciferase values were normalized according to renilla luciferase activity (n = 4; *p,0.001). doi:10.1371/journal.pone.0044651.g003
Techniques Used: Variant Assay, Transfection, Luciferase, Construct, Binding Assay, Plasmid Preparation, Sequencing, Activity Assay
Figure Legend Snippet: Figure 4. The override of ROD1 down-modulation by hypoxia increased apoptosis. HEK-293 were transfected with plasmids encoding ROD1 (pCMV-ROD1) or with vector alone (pCMV) and the next day were exposed to 1% hypoxia for the indicated time. A) HEK- 293 growth curve. Data are expressed as % of T0 normoxic control. Significant differences between pCMV-ROD1 and pCMV transfected cells in the same experimental condition are indicated (n = 5–11; *p,0.001; #p,0.008). B) Cell death assessed by Trypan blue exclusion assay. Data are expressed as % of dead cells for each experimental point. Differences between hypoxic cells transfected with either pCMV- ROD1 or pCMV are statistically significant (n = 5–11; *p,0.001; #p,0.008). C) Transfected cells were exposed to hypoxia for 48 hrs and apoptosis was measured assessing the apoptotic fragmentation of cytoplasmic DNA (n = 3; *p,0.007; #p,0.03). doi:10.1371/journal.pone.0044651.g004
Techniques Used: Transfection, Plasmid Preparation, Control, Trypan Blue Exclusion Assay
Related Articles
Transfection:Article Title: ROD1 Is a Seedless Target Gene of Hypoxia-Induced miR-210 Article Snippet: Hypoxic conditions were maintained in defined atmosphere chambers (Billups-Rothenberg Inc.); cells were placed in the chamber and then a gas mixture of 5% CO 2 - 95% N 2 was injected for 20 min, yielding about 1% O 2 ; thereafter, the chamber was sealed and incubated at 37°C for the indicated time. .. Article Title: ROD1 is a seedless target gene of hypoxia-induced miR-210. Article Snippet: Hypoxic conditions were maintained in defined atmosphere chambers (Billups-Rothenberg Inc.); cells were placed in the chamber and then a gas mixture of 5% CO2 - 95% N2 was injected for 20 min, yielding about 1% O2 [18]; thereafter, the chamber was sealed and incubated at 37uC for the indicated time. .. |