sorcin sirna  (Santa Cruz Biotechnology)

Journal: Journal of Molecular Endocrinology

Article Title: Sorcin is involved during embryo implantation via activating VEGF/PI3K/Akt pathway in mice

doi: 10.1530/jme-17-0153

Figure Lengend Snippet: Figure 1: Expression of sorcin during the period of early pregnancy and during estrous cycle in mouse

Article Snippet: 128 129 2.4 In vivo sorcin knock-down 130 Pregnant female mice underwent mini laparotomy under anesthesia on D3 of pregnancy to 131 deliver 50 nM of sorcin siRNA (sc-41017; Santa Cruz, CA) in a final volume of 25 μl with 132 Lipofectamine® RNAiMAX reagent (Thermo Fisher Scientific) per manufacturer’s 133 instructions in one of the uterine horn.

Techniques: Expressing

Journal: Journal of Molecular Endocrinology

Article Title: Sorcin is involved during embryo implantation via activating VEGF/PI3K/Akt pathway in mice

doi: 10.1530/jme-17-0153

Figure Lengend Snippet: Figure 2: Immunohistochemistry of sorcin expression in mouse uteri during early pregnancy. Sorcin expression was measured in endometrium i.e. glandular epithelium (GE), luminal epithelium (LE), and stroma (S) at different days of pregnancy (D1, D4, D5, D6 and D7). Graph in lower panel shows image score analysis. IS, implantation site; NP, non-pregnant; NC, Negative control. p values are ap<0.001 vs.

Article Snippet: 128 129 2.4 In vivo sorcin knock-down 130 Pregnant female mice underwent mini laparotomy under anesthesia on D3 of pregnancy to 131 deliver 50 nM of sorcin siRNA (sc-41017; Santa Cruz, CA) in a final volume of 25 μl with 132 Lipofectamine® RNAiMAX reagent (Thermo Fisher Scientific) per manufacturer’s 133 instructions in one of the uterine horn.

Techniques: Immunohistochemistry, Expressing, Negative Control

Journal: Journal of Molecular Endocrinology

Article Title: Sorcin is involved during embryo implantation via activating VEGF/PI3K/Akt pathway in mice

doi: 10.1530/jme-17-0153

Figure Lengend Snippet: Figure 3: Effect of sorcin inhibition on the number of implantation sites. The right horn was injected with

Article Snippet: 128 129 2.4 In vivo sorcin knock-down 130 Pregnant female mice underwent mini laparotomy under anesthesia on D3 of pregnancy to 131 deliver 50 nM of sorcin siRNA (sc-41017; Santa Cruz, CA) in a final volume of 25 μl with 132 Lipofectamine® RNAiMAX reagent (Thermo Fisher Scientific) per manufacturer’s 133 instructions in one of the uterine horn.

Techniques: Inhibition, Injection

Journal: Journal of Molecular Endocrinology

Article Title: Sorcin is involved during embryo implantation via activating VEGF/PI3K/Akt pathway in mice

doi: 10.1530/jme-17-0153

Figure Lengend Snippet: Figure 5: Role of ovarian hormones in regulation of sorcin expression. (A) Ovariectomized mice treated with

Article Snippet: 128 129 2.4 In vivo sorcin knock-down 130 Pregnant female mice underwent mini laparotomy under anesthesia on D3 of pregnancy to 131 deliver 50 nM of sorcin siRNA (sc-41017; Santa Cruz, CA) in a final volume of 25 μl with 132 Lipofectamine® RNAiMAX reagent (Thermo Fisher Scientific) per manufacturer’s 133 instructions in one of the uterine horn.

Techniques: Expressing

Journal: Journal of Molecular Endocrinology

Article Title: Sorcin is involved during embryo implantation via activating VEGF/PI3K/Akt pathway in mice

doi: 10.1530/jme-17-0153

Figure Lengend Snippet: Figure 6: Effect of sorcin silencing on cytosolic free Ca+2 , VEGF level, VEGFR-2 expression in mouse

Article Snippet: 128 129 2.4 In vivo sorcin knock-down 130 Pregnant female mice underwent mini laparotomy under anesthesia on D3 of pregnancy to 131 deliver 50 nM of sorcin siRNA (sc-41017; Santa Cruz, CA) in a final volume of 25 μl with 132 Lipofectamine® RNAiMAX reagent (Thermo Fisher Scientific) per manufacturer’s 133 instructions in one of the uterine horn.

Techniques: Expressing

Journal: Journal of Molecular Endocrinology

Article Title: Sorcin is involved during embryo implantation via activating VEGF/PI3K/Akt pathway in mice

doi: 10.1530/jme-17-0153

Figure Lengend Snippet: Figure 7: The proliferation, migration and invasion assay of HUVEC cells. (A) The proliferation of HUVECs was detected in various groups; HUVECs were cultured with supernatants (20% V/V) from mouse primary EECs transfected with sorcin siRNA or scrambled siRNA. In additional group, a blocking anti-VEGF antibody was added in the supernatant from scrambled siRNA treated EECs. Details have been given in “materials and

Article Snippet: 128 129 2.4 In vivo sorcin knock-down 130 Pregnant female mice underwent mini laparotomy under anesthesia on D3 of pregnancy to 131 deliver 50 nM of sorcin siRNA (sc-41017; Santa Cruz, CA) in a final volume of 25 μl with 132 Lipofectamine® RNAiMAX reagent (Thermo Fisher Scientific) per manufacturer’s 133 instructions in one of the uterine horn.

Techniques: Migration, Invasion Assay, Cell Culture, Transfection, Blocking Assay

Journal: Journal of Molecular Endocrinology

Article Title: Sorcin is involved during embryo implantation via activating VEGF/PI3K/Akt pathway in mice

doi: 10.1530/jme-17-0153

Figure Lengend Snippet: Figure 8: Schematic hypothetical representation of the molecular mechanism of sorcin in endometrium

Article Snippet: 128 129 2.4 In vivo sorcin knock-down 130 Pregnant female mice underwent mini laparotomy under anesthesia on D3 of pregnancy to 131 deliver 50 nM of sorcin siRNA (sc-41017; Santa Cruz, CA) in a final volume of 25 μl with 132 Lipofectamine® RNAiMAX reagent (Thermo Fisher Scientific) per manufacturer’s 133 instructions in one of the uterine horn.

Techniques:

Journal: Diabetes

Article Title: Glucose-Induced Nuclear Shuttling of ChREBP Is Mediated by Sorcin and Ca 2+ Ions in Pancreatic β-Cells

doi: 10.2337/db10-1329

Figure Lengend Snippet: Sorcin silencing decreases glucose-stimulated insulin secretion in MIN6 β-cells. MIN6 β-cells were transfected with sorcin-specific or scrambled siRNAs before insulin secretion assay as described in . Data are means ± SEM from 3 separate experiments.

Article Snippet: To assess the effect of sorcin overexpression or silencing on endogenous ChREBP nuclear localization, dissociated islets were transduced with either sorcin (also expressing GFP) or GFP adenoviral particles as above or with shRNA sorcin or control lentiviral particles (Santa Cruz) according to the manufacturer’s instructions in 11 mmol/L glucose RPMI 1640 for 48 h followed by 1 h in 3 mmol/L glucose RPMI 1640 and 6 h in 17 mmol/L glucose RPMI 1640.

Techniques: Transfection

Journal: Diabetes

Article Title: Glucose-Induced Nuclear Shuttling of ChREBP Is Mediated by Sorcin and Ca 2+ Ions in Pancreatic β-Cells

doi: 10.2337/db10-1329

Figure Lengend Snippet: ChREBP-EGFP nuclear translocation evoked by [Ca 2+ ] cyt requires sorcin for coupling of the events. A - F : Live-cell images of ChREBP-EGFP in MIN6 β-cells (typical of n ≥7–10 cells) expressed either alone or in combination with sorcin siRNA ( F ; n >10). All cells starved (3 mmol/L glucose) overnight and were imaged while perifusing with indicated buffers in warm KRBH. In C , cells were preincubated (10 min) with nifedipine (nif; 10 μmol/L) and diazoxide (diaz; 250 μmol/L) before imaging. Unless specified as Ca 2+ -free (containing EGTA), all buffers contained 1.5 mmol/L Ca 2+ . Scale bar = 10 μm. G - I (upper panels): [Ca 2+ ] cyt signals evoked by glucose ( G , I ) and ATP ( H , I ). Traces show the responses from ≥30 cells (≥55 for sorcin-silenced cells), taken from at least 3 independent experiments (means ± SEM). G - I (bottom panels): Real-time quantification of ChREBP-EGFP (gray values) in nuclear and cytosolic compartments. Traces (scale reversed to time-match events in the upper panel) show average from ≥10 cells ( I ; ≥30) during various treatments, from 3 independent experiments (means ± SEM). nuc, nucleus; cyto, cytosol; glu, glucose. ** P < 0.005, *** P < 0.0005 for the effects of treatments. (A high-quality digital representation of this figure is available in the online issue.)

Article Snippet: To assess the effect of sorcin overexpression or silencing on endogenous ChREBP nuclear localization, dissociated islets were transduced with either sorcin (also expressing GFP) or GFP adenoviral particles as above or with shRNA sorcin or control lentiviral particles (Santa Cruz) according to the manufacturer’s instructions in 11 mmol/L glucose RPMI 1640 for 48 h followed by 1 h in 3 mmol/L glucose RPMI 1640 and 6 h in 17 mmol/L glucose RPMI 1640.

Techniques: Translocation Assay, Imaging

Journal: Diabetes

Article Title: Glucose-Induced Nuclear Shuttling of ChREBP Is Mediated by Sorcin and Ca 2+ Ions in Pancreatic β-Cells

doi: 10.2337/db10-1329

Figure Lengend Snippet: Sorcin colocalizes with ChREBP in MIN6 β-cells and controls its subcellular localization. A and B : MIN6 β-cells were transfected with c- myc –ChREBP alone ( A ) or in combination with sorcin-HA ( B ) and cultured in either 3 or 30 mmol/L glucose for 6 h as indicated. Immunocytochemistry and confocal imaging were performed as described in . Scale bars = 5 μm. C : Quantification of c- myc –ChREBP in the nucleus, for the 30 mmol/L glucose experiments presented in A and B . The intensities of c- myc staining were measured using Volocity 4.0 software, for both nuclear and cytoplasmic regions of interest, in each individual cell, and background intensities were subtracted. Nuclear ChREBP intensity is expressed as percentage of cytoplasmic intensity ( n ≥ 3 experiments, with 30 cells per condition). D : MIN6 β-cells were transfected with sorcin-HA in 25 mmol/L Dulbecco’s modified Eagle’s medium for 48 h before culturing in 3 mmol/L glucose for 16 h, followed by 3 or 30 mmol/L glucose for 6 h. The nuclear and cytosolic fractions were extracted, and Western blots were performed as described in . The blots were probed with anti-ChREBP ( top ), anti-HA ( middle ), or anti–α-tubulin antibodies ( bottom ). The experiment was repeated twice with similar results. E : MIN6 β-cells were cotransfected with either −148L-PK luci, −183L-PK luci, or −1081TXNIP luci together with GFP or sorcin as indicated before cell lysis and luciferase assays as described in ( n = 3 to 8). GFP, green fluorescent protein. (A high-quality digital representation of this figure is available in the online issue.)

Article Snippet: To assess the effect of sorcin overexpression or silencing on endogenous ChREBP nuclear localization, dissociated islets were transduced with either sorcin (also expressing GFP) or GFP adenoviral particles as above or with shRNA sorcin or control lentiviral particles (Santa Cruz) according to the manufacturer’s instructions in 11 mmol/L glucose RPMI 1640 for 48 h followed by 1 h in 3 mmol/L glucose RPMI 1640 and 6 h in 17 mmol/L glucose RPMI 1640.

Techniques: Transfection, Cell Culture, Immunocytochemistry, Imaging, Staining, Software, Modification, Western Blot, Lysis, Luciferase

Journal: Diabetes

Article Title: Glucose-Induced Nuclear Shuttling of ChREBP Is Mediated by Sorcin and Ca 2+ Ions in Pancreatic β-Cells

doi: 10.2337/db10-1329

Figure Lengend Snippet: Sorcin controls ChREBP subcellular location in primary mouse β-cells. A : Dissociated mouse islets were transduced for 48 h with null or sorcin-overexpressing adenoviruses in 11 mmol/L glucose RPMI 1640, and then incubated 1 h in 3 mmol/L glucose followed by 6 h in 17 mmol/L RPMI 1640, before immunochemistry and confocal imaging as described in . Scale bars = 5 μm. C : Dissociated mouse islets were transduced with control or sorcin-shRNA lentiviruses for 72 h in 11 mmol/L RPMI 1640 and then incubated for 6 h in 3 mmol/L RPMI 1640 before immunochemistry and confocal imaging as above. B and C : Quantification of endogenous ChREBP in the nucleus, for the experiments presented in A and C . The intensities of ChREBP staining were measured using Volocity 4.0 software, for both nuclear and cytoplasmic regions of interest, in each individual cell, and background intensities were subtracted. Nuclear ChREBP intensity is expressed as percentage of cytoplasmic intensity ( n = 3 experiments, with 24 to 29 cells per condition). (A high-quality digital representation of this figure is available in the online issue.)

Article Snippet: To assess the effect of sorcin overexpression or silencing on endogenous ChREBP nuclear localization, dissociated islets were transduced with either sorcin (also expressing GFP) or GFP adenoviral particles as above or with shRNA sorcin or control lentiviral particles (Santa Cruz) according to the manufacturer’s instructions in 11 mmol/L glucose RPMI 1640 for 48 h followed by 1 h in 3 mmol/L glucose RPMI 1640 and 6 h in 17 mmol/L glucose RPMI 1640.

Techniques: Incubation, Imaging, Transduction, Control, shRNA, Staining, Software

Journal: Diabetes

Article Title: Glucose-Induced Nuclear Shuttling of ChREBP Is Mediated by Sorcin and Ca 2+ Ions in Pancreatic β-Cells

doi: 10.2337/db10-1329

Figure Lengend Snippet: Proposed model of ChREBP activation via sorcin and Ca 2+ ions. High glucose and subsequent Ca 2+ -induced Ca 2+ release (CICR) cause ChREBP and sorcin dissociation in the cytosol. Activated ChREBP then translocates into the nucleus where it activates transcription. IP3R, inositol 1,4,5-trisphosphate receptors; ER, endoplasmic reticulum; RyR, ryanodine receptors; PLC, phospholipase C; VGCC, voltage-gated calcium channels; Nuc, nucleus. (A high-quality color representation of this figure is available in the online issue.)

Article Snippet: To assess the effect of sorcin overexpression or silencing on endogenous ChREBP nuclear localization, dissociated islets were transduced with either sorcin (also expressing GFP) or GFP adenoviral particles as above or with shRNA sorcin or control lentiviral particles (Santa Cruz) according to the manufacturer’s instructions in 11 mmol/L glucose RPMI 1640 for 48 h followed by 1 h in 3 mmol/L glucose RPMI 1640 and 6 h in 17 mmol/L glucose RPMI 1640.

Techniques: Activation Assay

Journal: Diabetes

Article Title: Glucose-Induced Nuclear Shuttling of ChREBP Is Mediated by Sorcin and Ca 2+ Ions in Pancreatic β-Cells

doi: 10.2337/db10-1329

Figure Lengend Snippet: Sorcin controls ChREBP subcellular location in primary mouse β-cells. A : Dissociated mouse islets were transduced for 48 h with null or sorcin-overexpressing adenoviruses in 11 mmol/L glucose RPMI 1640, and then incubated 1 h in 3 mmol/L glucose followed by 6 h in 17 mmol/L RPMI 1640, before immunochemistry and confocal imaging as described in . Scale bars = 5 μm. C : Dissociated mouse islets were transduced with control or sorcin-shRNA lentiviruses for 72 h in 11 mmol/L RPMI 1640 and then incubated for 6 h in 3 mmol/L RPMI 1640 before immunochemistry and confocal imaging as above. B and C : Quantification of endogenous ChREBP in the nucleus, for the experiments presented in A and C . The intensities of ChREBP staining were measured using Volocity 4.0 software, for both nuclear and cytoplasmic regions of interest, in each individual cell, and background intensities were subtracted. Nuclear ChREBP intensity is expressed as percentage of cytoplasmic intensity ( n = 3 experiments, with 24 to 29 cells per condition). (A high-quality digital representation of this figure is available in the online issue.)

Article Snippet: Adenovirus production was then performed as in Ref. . Sorcin (sc-41017-v) and control (sc-108080) short hairpin RNA (shRNA) mouse lentiviruses were purchased from Santa Cruz.

Techniques: Incubation, Imaging, Transduction, Control, shRNA, Staining, Software