Journal: The Journal of Neuroscience

Article Title: Titration of Syntaxin1 in Mammalian Synapses Reveals Multiple Roles in Vesicle Docking, Priming, and Release Probability

doi: 10.1523/JNEUROSCI.0187-13.2013

Figure Lengend Snippet: Hypomorphic expression of a Stx1Byfp fusion knock-in allele. A, Cloning strategy for the Stx1Byfp fusion allele. Double asterisks denote a modified exon 7 described in Gerber et. al. (2008). B, Diagrams depicting the orientations of plasma membrane-anchored wild-type (top) and EYFP-fused Stx1B (bottom). C, RT-PCR amplification of a full ∼1.6 kb transcript encoded by the Stx1Byfp allele from Stx1B+/yfp mRNA. D, Immunoblot showing severely reduced levels of Stx1B–YFP in Stx1Byfp/yfp;Stx1A−/− embryonic brain lysates compared with lysates from control littermates. β-tubulin was used as a loading control. E, Immunofluorescent staining of hippocampal mass cultures for extracellularly localized YFP. Stx1Byfp/yfp, but not wild-type controls transduced with intracellular fluorescent protein, is positive for surface expression of YFP. Scale bar, 10 μm. F, Peroxidase immunostaining of Stx1B–YFP fusion protein in sagittal sections of P14 hippocampus and cerebellum. Antibodies to synaptotagmin 1 (Syn1) and Rab3A were used as positive controls, while PBS corresponds to negative control. DG, Dentate gyrus; MF, hippocampal mossy fiber synapse. G, Immunoblot showing expression levels of SNARE proteins, Munc18-1, and other synaptic proteins in wild-type, Stx1Byfp/yfp, and Stx1A−/− P14 mice. Valosin-containing protein (VCP) was used as a loading control. Asterisks mark proteins with significant changes in expression levels.

Article Snippet: For RNAi knock-down experiments, a Stx1B -specific short hairpin RNA (shRNA) target sequence (5′-GAT CCC AGG CAC AAT GAG ATC ATC AAA-3′) was obtained using Genscript siRNA Target Finder ( http://www.genscript.com ).

Techniques: Expressing, Knock-In, Cloning, Modification, Clinical Proteomics, Membrane, Reverse Transcription Polymerase Chain Reaction, Amplification, Western Blot, Control, Staining, Transduction, Immunostaining, Negative Control

Journal: The Journal of Neuroscience

Article Title: Titration of Syntaxin1 in Mammalian Synapses Reveals Multiple Roles in Vesicle Docking, Priming, and Release Probability

doi: 10.1523/JNEUROSCI.0187-13.2013

Figure Lengend Snippet: Stx1Byfp/yfp;Stx1A−/− double mutants show premature lethality, gross morphological defects, and diminished Ca2+-evoked and spontaneous release. A, Weight curves for the Stx1Byfp mouse line. Stx1A+/+;Stx1Byfp/yfp and StxA+/−; Stx1Byfp/yfp show impaired growth and development before death at 3 weeks. B, Survival curves for the Stx1Byfp mouse line. Stx1A+/+;Stx1Byfp/yfp and Stx1A+/−;Stx1Byfp/yfp die postnatally after 3 weeks. Stx1A−/−;Stx1Byfp/yfp are not shown because they die at birth. C, Gross morphological abnormalities in Stx1Byfp/yfp;Stx1A−/− embryos compared with Stx1B+/yfp;Stx1A−/− and Stx1B+/+;Stx1A−/− littermates at E18.5. D, Sample traces of EPSCs from Stx1B+/+;Stx1A−/− (black), Stx1B+/yfp;Stx1A−/− (blue), and Stx1Byfp/yfp;Stx1A−/− (red) autaptic neurons. E, Normalized summary plot of EPSC peak amplitudes (***p < 0.0001). F, Normalized summary plot of EPSC rise times (***p < 0.0001). G, Sample traces of mEPSCs recorded at −70 mV from Stx1B+/+;Stx1A−/− (black) and Stx1Byfp/yfp;Stx1A−/− (red) neurons. H, Normalized summary plot of mEPSC frequency (***p < 0.0001).

Article Snippet: For RNAi knock-down experiments, a Stx1B -specific short hairpin RNA (shRNA) target sequence (5′-GAT CCC AGG CAC AAT GAG ATC ATC AAA-3′) was obtained using Genscript siRNA Target Finder ( http://www.genscript.com ).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Titration of Syntaxin1 in Mammalian Synapses Reveals Multiple Roles in Vesicle Docking, Priming, and Release Probability

doi: 10.1523/JNEUROSCI.0187-13.2013

Figure Lengend Snippet: Stx1Byfp/yfp;Stx1A−/− double mutants have a smaller RRP of vesicles and slower RRP refilling rate. A, Sample traces of release induced by hyperosmotic stimulation from Stx1B+/+;Stx1A−/− (black), Stx1B+/yfp;Stx1A−/− (blue), and Stx1Byfp/yfp;Stx1A−/− (red) autaptic neurons. B, Normalized summary plot of RRP charge (***p < 0.0001). C, Refilling of the RRP as measured by paired applications of 0.5 m sucrose in Stx1B+/+;Stx1A−/− (black) and Stx1Byfp/yfp;Stx1A−/− (red). D, Fraction of the RRP refilled 2, 5, and 10 s after the first hyperosmotic stimulus (**p < 0.001).

Article Snippet: For RNAi knock-down experiments, a Stx1B -specific short hairpin RNA (shRNA) target sequence (5′-GAT CCC AGG CAC AAT GAG ATC ATC AAA-3′) was obtained using Genscript siRNA Target Finder ( http://www.genscript.com ).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Titration of Syntaxin1 in Mammalian Synapses Reveals Multiple Roles in Vesicle Docking, Priming, and Release Probability

doi: 10.1523/JNEUROSCI.0187-13.2013

Figure Lengend Snippet: Vesicle fusogenicity is impaired in Stx1Byfp/yfp;Stx1A−/− neurons. A, Sample traces of evoked responses from Stx1B+/+;Stx1A−/− (black) and Stx1Byfp/yfp;Stx1A−/− (red) neurons at different external Ca2+ concentrations. B, Ca2+ dose–response curves for Stx1B+/+;Stx1A−/− (black) and Stx1Byfp/yfp;Stx1A−/− (red) neurons. C, Normalized summary plot of Pvr, measured as the ratio of the EPSC charge and RRP charge, in Stx1B+/+;Stx1A−/− (black), Stx1B+/yfp;Stx1A−/− (blue), and Stx1Byfp/yfp;Stx1A−/− (red) neurons (***p < 0.0001). D, Short-term plasticity behavior during 10 Hz stimulation. E, Sample traces of release induced by different (hyperosmotic) sucrose concentrations. F, Dose–response curves from hypertonic sucrose stimulation. G, Spontaneous release rate, calculated as the fraction of the RRP released spontaneously per second (*p < 0.05).

Article Snippet: For RNAi knock-down experiments, a Stx1B -specific short hairpin RNA (shRNA) target sequence (5′-GAT CCC AGG CAC AAT GAG ATC ATC AAA-3′) was obtained using Genscript siRNA Target Finder ( http://www.genscript.com ).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Titration of Syntaxin1 in Mammalian Synapses Reveals Multiple Roles in Vesicle Docking, Priming, and Release Probability

doi: 10.1523/JNEUROSCI.0187-13.2013

Figure Lengend Snippet: Presynaptic Ca2+ influx is not altered in Stx1Byfp/yfp;Stx1A−/− neurons. A, Example baseline fluorescence (left; average of three images; F0) and peak 5 AP fluorescence (right; average two images; ΔF) for an Stx1Byfp/yfp;Stx1A−/− autaptic neuron. B, Average ΔF/F0 for ROIs selected from neuron shown in A (n = 41) in response to 2, 3, and 5 AP at 20 Hz. Arrow indicates initiation of stimulation. C, Average ΔF/F0 responses per cell for Stx1Byfp/yfp;Stx1A−/− (red; n = 35 cells) and control (black; Stx1B+/+;Stx1A−/− and Stx1B+/yfp;Stx1A−/− pooled; n = 33) for 2, 3, 5, and 10 AP at 20 Hz. D, E, Data were fitted with a linear regression analysis (red and black lines, respectively). Average paired-pulse ratio (D; 50 ms interstimulus interval; ***p < 0.0001) and EPSC amplitude (E; ***p < 0.0001) for Stx1Byfp/yfp;Stx1A−/− and control cells from C. All error bars represent SEM.

Article Snippet: For RNAi knock-down experiments, a Stx1B -specific short hairpin RNA (shRNA) target sequence (5′-GAT CCC AGG CAC AAT GAG ATC ATC AAA-3′) was obtained using Genscript siRNA Target Finder ( http://www.genscript.com ).

Techniques: Fluorescence, Control

Journal: The Journal of Neuroscience

Article Title: Titration of Syntaxin1 in Mammalian Synapses Reveals Multiple Roles in Vesicle Docking, Priming, and Release Probability

doi: 10.1523/JNEUROSCI.0187-13.2013

Figure Lengend Snippet: Knockdown of Stx1B–YFP further impaired release efficiency. A, Immunoblot showing knockdown of both Stx1B (in wild-type background), and Stx1B–YFP fusion protein (in Stx1Byfp/yfp;Stx1A−/− background), but not Stx1A, using a Stx1B sequence-specific shRNA. B, Sample traces of EPSCs from Stx1B+/+;Stx1A−/− (black), Stx1Byfp/yfp;Stx1A−/− untransduced (red), transduced with lentiviral vector only (Lois et al., 2002), and transduced with knock-down construct (purple). C, Sample traces of RRP released by hyperosmotic stimulation. D, Normalized summary plot of EPSC amplitudes (***p < 0.0001). E, Normalized summary plot of RRP charge (*p < 0.05, ***p < 0.001). F, Normalized summary plot of vesicular release probability. G, Short-term plasticity behavior during 10 Hz stimulation.

Article Snippet: For RNAi knock-down experiments, a Stx1B -specific short hairpin RNA (shRNA) target sequence (5′-GAT CCC AGG CAC AAT GAG ATC ATC AAA-3′) was obtained using Genscript siRNA Target Finder ( http://www.genscript.com ).

Techniques: Knockdown, Western Blot, Sequencing, shRNA, Transduction, Plasmid Preparation, Construct

Journal: The Journal of Neuroscience

Article Title: Titration of Syntaxin1 in Mammalian Synapses Reveals Multiple Roles in Vesicle Docking, Priming, and Release Probability

doi: 10.1523/JNEUROSCI.0187-13.2013

Figure Lengend Snippet: Knockdown of wild-type Stx1B phenocopied Stx1Byfp/yfp;Stx1A−/− hypomorphs. A, Sample traces of EPSCs from Stx1B+/+;Stx1A−/− neurons untransduced (black), transduced with lentiviral vector only (blue), or transduced with a Stx1B sequence-specific shRNA (green). B, Normalized summary plot of EPSC amplitudes (***p < 0.0001). C, Sample traces of RRP released by hyperosmotic stimulation. D, Normalized summary plot of RRP charge (***p < 0.0001). E, Normalized summary plot of vesicular release probability (**p < 0.001). F, Short-term plasticity behavior during 10 Hz stimulation. G, Sample images of autaptic neurons double labeled with the synaptic marker VGluT1 and the dendritic marker MAP2. Scale bar, 10 μm. H, Summary plot of synaptic vesicle density measured as VGluT1 puncta per 100 μm of dendrite length.

Article Snippet: For RNAi knock-down experiments, a Stx1B -specific short hairpin RNA (shRNA) target sequence (5′-GAT CCC AGG CAC AAT GAG ATC ATC AAA-3′) was obtained using Genscript siRNA Target Finder ( http://www.genscript.com ).

Techniques: Knockdown, Transduction, Plasmid Preparation, Sequencing, shRNA, Labeling, Marker

Journal: The Journal of Neuroscience

Article Title: Titration of Syntaxin1 in Mammalian Synapses Reveals Multiple Roles in Vesicle Docking, Priming, and Release Probability

doi: 10.1523/JNEUROSCI.0187-13.2013

Figure Lengend Snippet: Lentiviral expression of either Stx1A or Stx1B, but not Munc18-1, rescues synaptic release in neurons with severe reduction in Stx1 levels. A–C, Normalized summary plots of EPSC peak amplitudes (A), RRP charge (B), and vesicular release probability (C) in control Stx1Byfp/yfp;Stx1A−/− neurons (black), and Stx1Byfp/yfp;Stx1A−/− neurons transduced without (red) or with Stx1A, Stx1B, or Munc18-1 rescue constructs (red outlines; ANOVA with Tukey's test, ***p < 0.001; n.s., not significant). D–F, Normalized summary plots of EPSC peak amplitudes (D), RRP charge (E), and vesicular release probability (F) in Stx1A−/− neurons without (black) or with shRNA-mediated knockdown of Stx1B (teal), and knockdown neurons transduced with Stx1A, Stx1B, or Munc18-1 rescue constructs (teal outlines; ANOVA with Tukey's test, *p < 0.05, ***p < 0.001; n.s., not significant).

Article Snippet: For RNAi knock-down experiments, a Stx1B -specific short hairpin RNA (shRNA) target sequence (5′-GAT CCC AGG CAC AAT GAG ATC ATC AAA-3′) was obtained using Genscript siRNA Target Finder ( http://www.genscript.com ).

Techniques: Expressing, Control, Construct, shRNA, Knockdown, Transduction

Journal: The Journal of Neuroscience

Article Title: Titration of Syntaxin1 in Mammalian Synapses Reveals Multiple Roles in Vesicle Docking, Priming, and Release Probability

doi: 10.1523/JNEUROSCI.0187-13.2013

Figure Lengend Snippet: Vesicle distance from the active zone upon reduction of Stx1 expression. A, Sample tomographic images of presynaptic nerve terminals from Stx1B+/+;Stx1A−/− neurons and Stx1Byfp/yfp;Stx1A−/− neurons that were untransduced and transduced with a Stx1B sequence-specific shRNA. Scale bar, 100 nm. B, Cumulative distribution of vesicle distance from the plasma membrane. C, Summary plot of vesicle distance within 40 nm of the plasma membrane. *p < 0.05. D, Summary plot of presynaptic area. E, Summary plot of total number of synaptic vesicles per synapse. **p < 0.001. F, Summary plot of synaptic vesicle density (ANOVA with Tukey's test, **p < 0.001). G, Summary plot of active zone length.

Article Snippet: For RNAi knock-down experiments, a Stx1B -specific short hairpin RNA (shRNA) target sequence (5′-GAT CCC AGG CAC AAT GAG ATC ATC AAA-3′) was obtained using Genscript siRNA Target Finder ( http://www.genscript.com ).

Techniques: Expressing, Transduction, Sequencing, shRNA, Clinical Proteomics, Membrane

Journal: The Journal of Neuroscience

Article Title: Titration of Syntaxin1 in Mammalian Synapses Reveals Multiple Roles in Vesicle Docking, Priming, and Release Probability

doi: 10.1523/JNEUROSCI.0187-13.2013

Figure Lengend Snippet: Priming and release probability as a function of Stx1 expression levels. A, Normalized summary plot of Stx1B fluorescence intensities. ***p < 0.0001. B, Normalized summary plot of Bassoon fluorescence intensities. C, Sample images from double labeling of autaptic neurons with anti-Stx1B and anti-Bassoon antibodies. Scale bar, 5 μm. D, Summary plot of Stx1B/Bassoon fluorescence intensity ratios measured within regions marked by Bassoon. E, F, Readily releasable pool size (E) and vesicular release probability (F), derived from previously shown data, were plotted as a function of normalized Stx1B expression levels (Stx1B+/+;Stx1A−/−, black; Stx1B+/yfp;Stx1A−/−, blue; Stx1B+/+;Stx1A−/− + k.d., teal; Stx1Byfp/yfp;Stx1A−/−, red; Stx1Byfp/yfp;Stx1A−/− + k.d., purple).

Article Snippet: For RNAi knock-down experiments, a Stx1B -specific short hairpin RNA (shRNA) target sequence (5′-GAT CCC AGG CAC AAT GAG ATC ATC AAA-3′) was obtained using Genscript siRNA Target Finder ( http://www.genscript.com ).

Techniques: Expressing, Fluorescence, Labeling, Derivative Assay

Journal: Biology Open

Article Title: Lhx1 in the proximal region of the optic vesicle permits neural retina development in the chicken

doi: 10.1242/bio.20121396

Figure Lengend Snippet: Whole mount in situ hybridization (WISH) of embryos at Hamburger's and Hamilton's stages 9 ( A , B ), stage 10 ( C , D ), stage 11 ( E ), and stage 13 ( J ). (A,C,E) Dorsal views. (B,D,J) Lateral views. Transverse sections cut after WISH are shown in ( E-a , E-m , E-p ). At the middle OV level (E-m), Lhx1 is distinctly expressed in the proximal region of the OV and dorsal diencephalon (di). Outside these regions, Lhx1 is also expressed in the anterior neural ridge (anr) and notochord (nc). ( F , L ) Immunohistochemistry of Lhx1/Lhx5 protein. Transverse sections at the OV level of stage 11 (F), and stage 13 (L). ( G–I ) Expression patterns of Wnt8b (G), Pax2 (H), and Rx1 (I) at stage 11. Transverse sections at the OV level are shown for comparison. ( J – L ) At stage 13, Lhx1 is expressed in the dorsal diencephalon and by a subset of cell in the ventral diencephalon (arrow in J–L), but not in the presumptive neural retina (arrowhead in L). Scale bars: 100 µm.

Article Snippet: The Lhx1 or Lhx5 short hairpin RNA (shRNA) target sequences were designed by GenScript ( http://www.genscript.com ) ( ) and constructed according to Das et al. ( ).

Techniques: In Situ Hybridization, Immunohistochemistry, Expressing, Comparison

Journal: Biology Open

Article Title: Lhx1 in the proximal region of the optic vesicle permits neural retina development in the chicken

doi: 10.1242/bio.20121396

Figure Lengend Snippet: ( A ) Phylogenetic tree of chicken LIM-homeodomain (LIM-HD) factors. The LIM-HD factors are depicted by the NJ (neighbor-joining) method. Chicken LMO3 was used as an outgroup. The scale bar is calibrated in substitutions per site. Numbers show bootstrap confidence values. Amino acid sequences used in the tree construction were deduced from the nucleotide sequences listed in supplementary material Table S2 . ( B–D ) Lhx2 is expressed in the forebrain and whole OV at stage 11. Dorsal (B) and lateral (C) views. (D) Transverse section of the OV. ( E–H ) EGFP fluorescence images at 24 hours post-electroporation. Overexpression of Lhx2 (F) did not induce transformation of the outer optic cup to the NR, whereas overexpression of Lhx5 (G) or Lhx1 (H) induced a protrusion of the OV. ( I , J ) Histology of an Lhx5 -overexpressing retina at stage 24. A second NR transformed from the outer layer of the optic cup. ( K , L ) In situ hybridization of an Lhx5 -overexpressing retina at stage 21. Arrows show activation of Chx10 (K) and suppression of Otx2 (L) in the developing second NR. ( M–Q ) Whole mount in situ hybridization (WISH) of embryos at Hamburger's and Hamilton's stages 9 (M,N), stage 10 (O,P), and stage 11 (Q). (M,O,Q) Dorsal views. (N,P) Lateral views. Transverse sections cut after WISH are shown in ( Q-a , Q-m , Q-p ). At the middle OV level (Q-m), Lhx5 is distinctly expressed in the proximal region of the OV, while it is expressed in the ventral diencephalon at the posterior OV level (Q-p). ( R–T ) EGFP fluorescence (R) and in situ hybridization of Lhx1 (S,T) at 24 hours post-electroporation. Lhx1 expression is ectopically induced in the protruding outer layer of the optic cup after Lhx5 overexpression (arrowhead in S,T). Scale bars: 100 µm.

Article Snippet: The Lhx1 or Lhx5 short hairpin RNA (shRNA) target sequences were designed by GenScript ( http://www.genscript.com ) ( ) and constructed according to Das et al. ( ).

Techniques: Fluorescence, Electroporation, Over Expression, Transformation Assay, In Situ Hybridization, Activation Assay, Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: ERβ compensates for the absence of ERα function to promote osteoblast viability by inhibition of SOST signaling

doi: 10.3892/etm.2017.5014

Figure Lengend Snippet: ERα and ERβ knockdown in MCE cells. Western blot analysis was performed for ERα and ERβ after shRNA-ERα or shRNA-ERβ treatment. **P<0.01, ***P<0.001 vs. NC. ER, estrogen receptor; shRNA, short hairpin RNA; NC, normal control.

Article Snippet: ERα short hairpin RNA (shRNA) sequences: (CCG GTA CAG GCC AAA TTC AGA TAA CTC GAG TTA TCT GAA TTT GGC CTG TAG TTT TT) and ERβ shRNA (CCG GGC GAG TAA CAA GGG CAT GGA ACT CGA GTT CCA TGC CCT TGT TAC TCG CTT TTT) sequences were designed and synthesized by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Knockdown, Western Blot, shRNA, Control

Journal: Experimental and Therapeutic Medicine

Article Title: ERβ compensates for the absence of ERα function to promote osteoblast viability by inhibition of SOST signaling

doi: 10.3892/etm.2017.5014

Figure Lengend Snippet: Effect of ERα and ERβ on MCE cell cycle. Cell cycle analysis in MCE cells treated with (A) shRNA-ERα alone, (B) co-treated with shRNA-ERα and shRNA-ERβ or (C) co-treated with shRNA-ERα and ERβ agonist DPN. *P<0.05, **P<0.01 as indicated. ER, estrogen receptor; shRNA, short hairpin RNA; NC, normal control; DPN, 2,3-bis(4-hydroxyphenyl)-propionitrile.

Article Snippet: ERα short hairpin RNA (shRNA) sequences: (CCG GTA CAG GCC AAA TTC AGA TAA CTC GAG TTA TCT GAA TTT GGC CTG TAG TTT TT) and ERβ shRNA (CCG GGC GAG TAA CAA GGG CAT GGA ACT CGA GTT CCA TGC CCT TGT TAC TCG CTT TTT) sequences were designed and synthesized by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Cell Cycle Assay, shRNA, Control

Journal: Experimental and Therapeutic Medicine

Article Title: ERβ compensates for the absence of ERα function to promote osteoblast viability by inhibition of SOST signaling

doi: 10.3892/etm.2017.5014

Figure Lengend Snippet: Effect of ERα and ERβ on MCE cell viability. MTT assay performed in cells treated with shRNA-ERα alone or co-treated with shRNA-ERα and shRNA-ERβ or co-treated with shRNA-ERα and ERβ agonist DPN. *P<0.05, shRNA-ERα vs. shRNA-ERα + shRNA-ERβ; # P<0.05, shRNA-ERα vs. shRNA-ERα+DPN; $ P<0.05, shRNA-ERα vs. NC. OD, optical density; ER, estrogen receptor; shRNA, short hairpin RNA; NC, normal control; DPN, 2,3-bis(4-hydroxyphenyl)-propionitrile.

Article Snippet: ERα short hairpin RNA (shRNA) sequences: (CCG GTA CAG GCC AAA TTC AGA TAA CTC GAG TTA TCT GAA TTT GGC CTG TAG TTT TT) and ERβ shRNA (CCG GGC GAG TAA CAA GGG CAT GGA ACT CGA GTT CCA TGC CCT TGT TAC TCG CTT TTT) sequences were designed and synthesized by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: MTT Assay, shRNA, Control

Journal: Experimental and Therapeutic Medicine

Article Title: ERβ compensates for the absence of ERα function to promote osteoblast viability by inhibition of SOST signaling

doi: 10.3892/etm.2017.5014

Figure Lengend Snippet: Effect of ERα and ERβ on SOST, Runx2 and OPG expression. Reverse transcription-quantitative polymerase chain reaction analysis for (A) SOST, (B) Runx2 and (C) OPG in cells with shRNA-ERα alone or co-treated with shRNA-ERα and shRNA-ERβ or co-treated with shRNA-ERα and ERβ agonist DPN. *P<0.05, **P<0.01, ***P<0.001 as indicated. ER, estrogen receptor; shRNA, short hairpin RNA; NC, normal control; DPN, 2,3-bis(4-hydroxyphenyl)-propionitrile.

Article Snippet: ERα short hairpin RNA (shRNA) sequences: (CCG GTA CAG GCC AAA TTC AGA TAA CTC GAG TTA TCT GAA TTT GGC CTG TAG TTT TT) and ERβ shRNA (CCG GGC GAG TAA CAA GGG CAT GGA ACT CGA GTT CCA TGC CCT TGT TAC TCG CTT TTT) sequences were designed and synthesized by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, shRNA, Control

Journal: Experimental and Therapeutic Medicine

Article Title: ERβ compensates for the absence of ERα function to promote osteoblast viability by inhibition of SOST signaling

doi: 10.3892/etm.2017.5014

Figure Lengend Snippet: Effect of ERα and ERβ on Runx2 and OPG protein levels. ELISA was conducted for protein concentrations of (A) Runx2 and (B) OPG in cells with shRNA-ERα alone or co-treated with shRNA-ERα and shRNA-ERβ or co-treated with shRNA-ERα and DPN or co-treated with shRNA-ERα, shRNA-ERβ and Sost antibody. *P<0.05, **P<0.01, ***P<0.001 as indicated. ER, estrogen receptor; shRNA, short hairpin RNA; NC, normal control; DPN, 2,3-bis(4-hydroxyphenyl)-propionitrile.

Article Snippet: ERα short hairpin RNA (shRNA) sequences: (CCG GTA CAG GCC AAA TTC AGA TAA CTC GAG TTA TCT GAA TTT GGC CTG TAG TTT TT) and ERβ shRNA (CCG GGC GAG TAA CAA GGG CAT GGA ACT CGA GTT CCA TGC CCT TGT TAC TCG CTT TTT) sequences were designed and synthesized by Guangzhou RiboBio Co., Ltd. (Guangzhou, China).

Techniques: Enzyme-linked Immunosorbent Assay, shRNA, Control