Review

hap1 overexpression (OriGene)

OriGene is a verified supplier

OriGene manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

OriGene hap1 overexpression

Analysis of mRNA that encode store-operated calcium entry (SOCE) proteins in day in vitro 21 (DIV21) medium spiny neuron (MSN) cultures from YAC128 and wildtype mice. (A) Gene expression analysis of mRNA that encode SOCE proteins and proteins that are involved in Ca 2+ homeostasis in MSNs from wildtype (control) and YAC128 mice using real-time polymerase chain reaction (PCR). The gene expression results for all genes were normalized to Gapdh . Data from wildtype MSNs were normalized to 1. (B) Relative mRNA levels of genes that encode SOCE proteins, measured as 2 ∧ -dCt. (C) Immunoblots of HTT-associated protein-1 <t>(HAP1)</t> normalized to GAPDH and CacyBP/SIP (both monomers and dimers) normalized to Vinculin in YAC128 and wildtype MSN cultures. On the bottom of each panel, the expression levels of <t>HAP1</t> and CacyBP/SIP proteins are plotted. The results are expressed as mean ± SEM and represent data from three independent mRNA and protein preparations of three different MSN cultures. ns, not significant.

Hap1 Overexpression, supplied by OriGene, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/hap1+overexpression/pmc06231533-77-34-11?v=OriGene
Average 91 stars, based on 3 article reviews

hap1 overexpression - by Bioz Stars, 2026-08

91/100 stars

Images

1) Product Images from "Huntingtin-Associated Protein 1A Regulates Store-Operated Calcium Entry in Medium Spiny Neurons From Transgenic YAC128 Mice, a Model of Huntington’s Disease"

Article Title: Huntingtin-Associated Protein 1A Regulates Store-Operated Calcium Entry in Medium Spiny Neurons From Transgenic YAC128 Mice, a Model of Huntington’s Disease

Journal: Frontiers in Cellular Neuroscience

doi: 10.3389/fncel.2018.00381

Figure Legend Snippet: Analysis of mRNA that encode store-operated calcium entry (SOCE) proteins in day in vitro 21 (DIV21) medium spiny neuron (MSN) cultures from YAC128 and wildtype mice. (A) Gene expression analysis of mRNA that encode SOCE proteins and proteins that are involved in Ca 2+ homeostasis in MSNs from wildtype (control) and YAC128 mice using real-time polymerase chain reaction (PCR). The gene expression results for all genes were normalized to Gapdh . Data from wildtype MSNs were normalized to 1. (B) Relative mRNA levels of genes that encode SOCE proteins, measured as 2 ∧ -dCt. (C) Immunoblots of HTT-associated protein-1 (HAP1) normalized to GAPDH and CacyBP/SIP (both monomers and dimers) normalized to Vinculin in YAC128 and wildtype MSN cultures. On the bottom of each panel, the expression levels of HAP1 and CacyBP/SIP proteins are plotted. The results are expressed as mean ± SEM and represent data from three independent mRNA and protein preparations of three different MSN cultures. ns, not significant.

Techniques Used: In Vitro, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Parameters of Ca 2+ homeostasis in YAC128 MSNs that overexpress HAP1A. (A) Immunoblots of HAP1 and green fluorescent protein (GFP) in YAC128 MSN cultures that overexpressed HAP1A-pLenti-GFP, HAP1B-pLenti-GFP and pLenti-GFP. (B) GFP fluorescence that indicates the overexpression of HAP1A-pLenti-GFP and HAP1B-pLenti-GFP and pLenti-GFP in YAC128 MSNs. (C) Protocol to induce SOCE in YAC128 MSNs that overexpressed HAP1 isoforms using the eight-well system. (D) Protocol to induce SOCE in wildtype MSNs that overexpressed HAP1 isoforms. Cultures of MSNs on DIV14 from YAC128 and wildtype mice (control) that overexpressed HAP1A-pLenti-GFP, HAP1B-pLenti-GFP, or pLenti-GFP using lentiviruses were loaded with the Ca 2+ indicator Fura-2AM and incubated in Ca 2+ -free medium (0.1 mM EGTA). Ca 2+ release from the endoplasmic reticulum (ER) was induced by 20 μM DHPG. SOCE was activated by the addition of 2 mM Ca 2+ to the medium. KCl (56 nM) in 2 mM Ca 2+ was applied to distinguish neurons from glial cells. (E) Ca 2+ release from the ER in YAC128 MSNs that overexpressed HAP1 isoforms. (F) Ca 2+ release from the ER in wildtype MSNs that overexpressed HAP1 isoforms. (G) Ca 2+ influx via DHPG-induced SOCE in YAC128 MSNs that overexpressed HAP1 isoforms. (H) Ca 2+ influx via DHPG-induced SOCE in wildtype MSNs that overexpressed HAP1 isoforms. The results are expressed as mean ± SEM. The number of cells is shown on the top of the bars. The results of at least three independent MSN culture preparations are shown. * p < 0.05, *** p < 0.001. ns, not significant.

Figure Legend Snippet: Parameters of Ca 2+ homeostasis in YAC128 MSNs that overexpress HAP1A. (A) Immunoblots of HAP1 and green fluorescent protein (GFP) in YAC128 MSN cultures that overexpressed HAP1A-pLenti-GFP, HAP1B-pLenti-GFP and pLenti-GFP. (B) GFP fluorescence that indicates the overexpression of HAP1A-pLenti-GFP and HAP1B-pLenti-GFP and pLenti-GFP in YAC128 MSNs. (C) Protocol to induce SOCE in YAC128 MSNs that overexpressed HAP1 isoforms using the eight-well system. (D) Protocol to induce SOCE in wildtype MSNs that overexpressed HAP1 isoforms. Cultures of MSNs on DIV14 from YAC128 and wildtype mice (control) that overexpressed HAP1A-pLenti-GFP, HAP1B-pLenti-GFP, or pLenti-GFP using lentiviruses were loaded with the Ca 2+ indicator Fura-2AM and incubated in Ca 2+ -free medium (0.1 mM EGTA). Ca 2+ release from the endoplasmic reticulum (ER) was induced by 20 μM DHPG. SOCE was activated by the addition of 2 mM Ca 2+ to the medium. KCl (56 nM) in 2 mM Ca 2+ was applied to distinguish neurons from glial cells. (E) Ca 2+ release from the ER in YAC128 MSNs that overexpressed HAP1 isoforms. (F) Ca 2+ release from the ER in wildtype MSNs that overexpressed HAP1 isoforms. (G) Ca 2+ influx via DHPG-induced SOCE in YAC128 MSNs that overexpressed HAP1 isoforms. (H) Ca 2+ influx via DHPG-induced SOCE in wildtype MSNs that overexpressed HAP1 isoforms. The results are expressed as mean ± SEM. The number of cells is shown on the top of the bars. The results of at least three independent MSN culture preparations are shown. * p < 0.05, *** p < 0.001. ns, not significant.

Techniques Used: Western Blot, Fluorescence, Over Expression, Incubation

Parameters of Ca 2+ homeostasis in YAC128 MSNs with silencing of HAP1A. (A) Immunoblots of HAP1 and beta-actin in YAC128 MSN cultures that overexpressed short-hairpin RNA (shRNA) against HAP1. shRNAs against HAP1 are named a, b, c, d, control scrambled shRNA, and control MSN cultures as indicated on the blot. (B) Protocol to induce SOCE in YAC128 MSNs with silencing of HAP1 using the eight-well system. Cultures of MSNs on DIV14 from YAC128 mice that overexpressed different shRNAs against HAP1 in pLenti-GFP were loaded with the Ca 2+ indicator Fura-2AM and incubated in Ca 2+ -free medium (0.1 mM EGTA). Ca 2+ release from the ER was induced by 20 μM DHPG. SOCE was activated by the addition of 2 mM Ca 2+ to the medium. KCl (56 nM) in 2 mM Ca 2+ was applied to distinguish neurons from glial cells. (C) Ca 2+ release from the ER in YAC128 MSNs that overexpressed different shRNAs against HAP1. (D) Ca 2+ influx via DHPG-induced SOCE in YAC128 MSNs that overexpressed different shRNAs against HAP1. The results are expressed as mean ± SEM. The number of cells is shown on the top of the bars. The results of three independent MSN culture preparations are shown. *** p < 0.001.

Figure Legend Snippet: Parameters of Ca 2+ homeostasis in YAC128 MSNs with silencing of HAP1A. (A) Immunoblots of HAP1 and beta-actin in YAC128 MSN cultures that overexpressed short-hairpin RNA (shRNA) against HAP1. shRNAs against HAP1 are named a, b, c, d, control scrambled shRNA, and control MSN cultures as indicated on the blot. (B) Protocol to induce SOCE in YAC128 MSNs with silencing of HAP1 using the eight-well system. Cultures of MSNs on DIV14 from YAC128 mice that overexpressed different shRNAs against HAP1 in pLenti-GFP were loaded with the Ca 2+ indicator Fura-2AM and incubated in Ca 2+ -free medium (0.1 mM EGTA). Ca 2+ release from the ER was induced by 20 μM DHPG. SOCE was activated by the addition of 2 mM Ca 2+ to the medium. KCl (56 nM) in 2 mM Ca 2+ was applied to distinguish neurons from glial cells. (C) Ca 2+ release from the ER in YAC128 MSNs that overexpressed different shRNAs against HAP1. (D) Ca 2+ influx via DHPG-induced SOCE in YAC128 MSNs that overexpressed different shRNAs against HAP1. The results are expressed as mean ± SEM. The number of cells is shown on the top of the bars. The results of three independent MSN culture preparations are shown. *** p < 0.001.

Techniques Used: Western Blot, shRNA, Incubation

Effect of HAP1A on ER Ca 2+ content in YAC128 MSNs. Cultures of MSNs on DIV14 from YAC128 mice that overexpressed HAP1A-pLenti-GFP, HAP1B-pLenti-GFP, or pLenti-GFP using lentiviruses that were loaded with the Ca 2+ indicator Fura-2AM. (A) The figure shows the protocol to induce ionomycin-induced Ca 2+ release from the ER in YAC128 MSNs that overexpressed HAP1 isoforms using the eight-well system. YAC128 MSNs that overexpressed HAP1A isoforms were incubated in Ca 2 -free medium (0.1 mM EGTA), followed by 15 μM ionomycin application to induce the release of Ca 2+ . KCl (56 nM) in 2 mM Ca 2+ was applied to distinguish neurons from glial cells. (B) Ionomycin-induced ER Ca 2+ release in YAC128 MSNs that overexpressed HAP1 isoforms. The results are expressed as mean ± SEM. The number of cells is shown on the top of the bars. * p < 0.05. ns, not significant. The results were obtained from three independent MSN culture preparations.

Figure Legend Snippet: Effect of HAP1A on ER Ca 2+ content in YAC128 MSNs. Cultures of MSNs on DIV14 from YAC128 mice that overexpressed HAP1A-pLenti-GFP, HAP1B-pLenti-GFP, or pLenti-GFP using lentiviruses that were loaded with the Ca 2+ indicator Fura-2AM. (A) The figure shows the protocol to induce ionomycin-induced Ca 2+ release from the ER in YAC128 MSNs that overexpressed HAP1 isoforms using the eight-well system. YAC128 MSNs that overexpressed HAP1A isoforms were incubated in Ca 2 -free medium (0.1 mM EGTA), followed by 15 μM ionomycin application to induce the release of Ca 2+ . KCl (56 nM) in 2 mM Ca 2+ was applied to distinguish neurons from glial cells. (B) Ionomycin-induced ER Ca 2+ release in YAC128 MSNs that overexpressed HAP1 isoforms. The results are expressed as mean ± SEM. The number of cells is shown on the top of the bars. * p < 0.05. ns, not significant. The results were obtained from three independent MSN culture preparations.

Techniques Used: Incubation

Image Search Results

Journal: Frontiers in Cellular Neuroscience

Article Title: Huntingtin-Associated Protein 1A Regulates Store-Operated Calcium Entry in Medium Spiny Neurons From Transgenic YAC128 Mice, a Model of Huntington’s Disease

doi: 10.3389/fncel.2018.00381

Figure Lengend Snippet: Analysis of mRNA that encode store-operated calcium entry (SOCE) proteins in day in vitro 21 (DIV21) medium spiny neuron (MSN) cultures from YAC128 and wildtype mice. (A) Gene expression analysis of mRNA that encode SOCE proteins and proteins that are involved in Ca 2+ homeostasis in MSNs from wildtype (control) and YAC128 mice using real-time polymerase chain reaction (PCR). The gene expression results for all genes were normalized to Gapdh . Data from wildtype MSNs were normalized to 1. (B) Relative mRNA levels of genes that encode SOCE proteins, measured as 2 ∧ -dCt. (C) Immunoblots of HTT-associated protein-1 (HAP1) normalized to GAPDH and CacyBP/SIP (both monomers and dimers) normalized to Vinculin in YAC128 and wildtype MSN cultures. On the bottom of each panel, the expression levels of HAP1 and CacyBP/SIP proteins are plotted. The results are expressed as mean ± SEM and represent data from three independent mRNA and protein preparations of three different MSN cultures. ns, not significant.

Article Snippet: HAP1A or HAP1B mouse cDNA clones in pCMV6-ENTRY that originated from Origene (catalog no. NM010404 or NM177981) were cloned between the Sgf I and Mlu I sites into pLenti-C-mGFP (catalog no. PS100071, Origene) for HAP1 overexpression.

Techniques: In Vitro, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Frontiers in Cellular Neuroscience

Article Title: Huntingtin-Associated Protein 1A Regulates Store-Operated Calcium Entry in Medium Spiny Neurons From Transgenic YAC128 Mice, a Model of Huntington’s Disease

doi: 10.3389/fncel.2018.00381

Figure Lengend Snippet: Parameters of Ca 2+ homeostasis in YAC128 MSNs that overexpress HAP1A. (A) Immunoblots of HAP1 and green fluorescent protein (GFP) in YAC128 MSN cultures that overexpressed HAP1A-pLenti-GFP, HAP1B-pLenti-GFP and pLenti-GFP. (B) GFP fluorescence that indicates the overexpression of HAP1A-pLenti-GFP and HAP1B-pLenti-GFP and pLenti-GFP in YAC128 MSNs. (C) Protocol to induce SOCE in YAC128 MSNs that overexpressed HAP1 isoforms using the eight-well system. (D) Protocol to induce SOCE in wildtype MSNs that overexpressed HAP1 isoforms. Cultures of MSNs on DIV14 from YAC128 and wildtype mice (control) that overexpressed HAP1A-pLenti-GFP, HAP1B-pLenti-GFP, or pLenti-GFP using lentiviruses were loaded with the Ca 2+ indicator Fura-2AM and incubated in Ca 2+ -free medium (0.1 mM EGTA). Ca 2+ release from the endoplasmic reticulum (ER) was induced by 20 μM DHPG. SOCE was activated by the addition of 2 mM Ca 2+ to the medium. KCl (56 nM) in 2 mM Ca 2+ was applied to distinguish neurons from glial cells. (E) Ca 2+ release from the ER in YAC128 MSNs that overexpressed HAP1 isoforms. (F) Ca 2+ release from the ER in wildtype MSNs that overexpressed HAP1 isoforms. (G) Ca 2+ influx via DHPG-induced SOCE in YAC128 MSNs that overexpressed HAP1 isoforms. (H) Ca 2+ influx via DHPG-induced SOCE in wildtype MSNs that overexpressed HAP1 isoforms. The results are expressed as mean ± SEM. The number of cells is shown on the top of the bars. The results of at least three independent MSN culture preparations are shown. * p < 0.05, *** p < 0.001. ns, not significant.

Techniques: Western Blot, Fluorescence, Over Expression, Incubation

Journal: Frontiers in Cellular Neuroscience

Article Title: Huntingtin-Associated Protein 1A Regulates Store-Operated Calcium Entry in Medium Spiny Neurons From Transgenic YAC128 Mice, a Model of Huntington’s Disease

doi: 10.3389/fncel.2018.00381

Figure Lengend Snippet: Parameters of Ca 2+ homeostasis in YAC128 MSNs with silencing of HAP1A. (A) Immunoblots of HAP1 and beta-actin in YAC128 MSN cultures that overexpressed short-hairpin RNA (shRNA) against HAP1. shRNAs against HAP1 are named a, b, c, d, control scrambled shRNA, and control MSN cultures as indicated on the blot. (B) Protocol to induce SOCE in YAC128 MSNs with silencing of HAP1 using the eight-well system. Cultures of MSNs on DIV14 from YAC128 mice that overexpressed different shRNAs against HAP1 in pLenti-GFP were loaded with the Ca 2+ indicator Fura-2AM and incubated in Ca 2+ -free medium (0.1 mM EGTA). Ca 2+ release from the ER was induced by 20 μM DHPG. SOCE was activated by the addition of 2 mM Ca 2+ to the medium. KCl (56 nM) in 2 mM Ca 2+ was applied to distinguish neurons from glial cells. (C) Ca 2+ release from the ER in YAC128 MSNs that overexpressed different shRNAs against HAP1. (D) Ca 2+ influx via DHPG-induced SOCE in YAC128 MSNs that overexpressed different shRNAs against HAP1. The results are expressed as mean ± SEM. The number of cells is shown on the top of the bars. The results of three independent MSN culture preparations are shown. *** p < 0.001.

Techniques: Western Blot, shRNA, Incubation

Journal: Frontiers in Cellular Neuroscience

Article Title: Huntingtin-Associated Protein 1A Regulates Store-Operated Calcium Entry in Medium Spiny Neurons From Transgenic YAC128 Mice, a Model of Huntington’s Disease

doi: 10.3389/fncel.2018.00381

Figure Lengend Snippet: Effect of HAP1A on ER Ca 2+ content in YAC128 MSNs. Cultures of MSNs on DIV14 from YAC128 mice that overexpressed HAP1A-pLenti-GFP, HAP1B-pLenti-GFP, or pLenti-GFP using lentiviruses that were loaded with the Ca 2+ indicator Fura-2AM. (A) The figure shows the protocol to induce ionomycin-induced Ca 2+ release from the ER in YAC128 MSNs that overexpressed HAP1 isoforms using the eight-well system. YAC128 MSNs that overexpressed HAP1A isoforms were incubated in Ca 2 -free medium (0.1 mM EGTA), followed by 15 μM ionomycin application to induce the release of Ca 2+ . KCl (56 nM) in 2 mM Ca 2+ was applied to distinguish neurons from glial cells. (B) Ionomycin-induced ER Ca 2+ release in YAC128 MSNs that overexpressed HAP1 isoforms. The results are expressed as mean ± SEM. The number of cells is shown on the top of the bars. * p < 0.05. ns, not significant. The results were obtained from three independent MSN culture preparations.

Techniques: Incubation

Review

Structured Review

Images

1) Product Images from "Huntingtin-Associated Protein 1A Regulates Store-Operated Calcium Entry in Medium Spiny Neurons From Transgenic YAC128 Mice, a Model of Huntington’s Disease"

Similar Products

Image Search Results