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sirna targeting bi 1  (Santa Cruz Biotechnology)


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    Santa Cruz Biotechnology sirna targeting bi 1
    Expression levels of endogenous <t>BI-1,</t> IRE1α, XBP1 and CHOP post-HI. (A) Representative immunoblots showing protein expression levels in ipsilateral hemispheric tissue from 10-day-old neonatal rats. (B–E) Quantitative analysis of BI-1 (B), IRE1α (C), XBP1 (D) and CHOP (E) time-dependent expression after HI (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus 6 h, @ P <0.05 versus 24 h; n =4. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.
    Sirna Targeting Bi 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/bi+1+sirna/BI-1+siRNA/pmc06898997-329-7-11
    Average 92 stars, based on 1 article reviews
    sirna targeting bi 1 - by Bioz Stars, 2026-09
    92/100 stars

    Images

    1) Product Images from "Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model"

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    Journal: Disease Models & Mechanisms

    doi: 10.1242/dmm.040352

    Expression levels of endogenous BI-1, IRE1α, XBP1 and CHOP post-HI. (A) Representative immunoblots showing protein expression levels in ipsilateral hemispheric tissue from 10-day-old neonatal rats. (B–E) Quantitative analysis of BI-1 (B), IRE1α (C), XBP1 (D) and CHOP (E) time-dependent expression after HI (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus 6 h, @ P <0.05 versus 24 h; n =4. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.
    Figure Legend Snippet: Expression levels of endogenous BI-1, IRE1α, XBP1 and CHOP post-HI. (A) Representative immunoblots showing protein expression levels in ipsilateral hemispheric tissue from 10-day-old neonatal rats. (B–E) Quantitative analysis of BI-1 (B), IRE1α (C), XBP1 (D) and CHOP (E) time-dependent expression after HI (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus 6 h, @ P <0.05 versus 24 h; n =4. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Techniques Used: Expressing, Western Blot

    Ad-TMBIM6 administered at 48 h pre-HI reduced percentage infarcted area and showed localization to neurons and microglia at 72 h post-HI. (A) Representative images of TTC-stained sections and quantification of percentage infarcted area at 72 h post-HI in brain tissue from neonatal rats expressing BI-1 through adenoviral transduction with Ad-TMBIM6. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle, n =6. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (B–E) Representative microphotographs of double immunofluorescence staining of BI-1 (red) with neurons (NeuN, green) (B), IRE1α (red) with neurons (NeuN, green) (C), BI-1 (red) with astrocytes (GFAP, green) (D) and BI-1 (red) with microglia (Iba-1, green) (E) in the peri-infarcted area at 72 h post-HI. Blue, DAPI. n =1/group. Merged images show localization of BI-1 or IRE1α to neurons, astrocytes or microglia. There was minimal localization of BI-1 to astrocytes. Scale bars: 50 μm.
    Figure Legend Snippet: Ad-TMBIM6 administered at 48 h pre-HI reduced percentage infarcted area and showed localization to neurons and microglia at 72 h post-HI. (A) Representative images of TTC-stained sections and quantification of percentage infarcted area at 72 h post-HI in brain tissue from neonatal rats expressing BI-1 through adenoviral transduction with Ad-TMBIM6. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle, n =6. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (B–E) Representative microphotographs of double immunofluorescence staining of BI-1 (red) with neurons (NeuN, green) (B), IRE1α (red) with neurons (NeuN, green) (C), BI-1 (red) with astrocytes (GFAP, green) (D) and BI-1 (red) with microglia (Iba-1, green) (E) in the peri-infarcted area at 72 h post-HI. Blue, DAPI. n =1/group. Merged images show localization of BI-1 or IRE1α to neurons, astrocytes or microglia. There was minimal localization of BI-1 to astrocytes. Scale bars: 50 μm.

    Techniques Used: Staining, Expressing, Transduction, Double Immunofluorescence Staining

    Expression of BI-1 improved long-term neurological deficits at 4 weeks post-HI. (A,B) In rat pups expressing BI-1 through treatment with Ad-TMBIM6 , long-term motor, learning and memory were assessed via foot fault test (A) and water maze test (B) at 4 weeks post-HI. (C,D) Quantitative analysis of percentage of weight of ipsilateral and contralateral regions out of total brain weights (C) and percentage of tissue loss (D) at 4 weeks post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; n =6–9. One-way ANOVA followed by Tukey multiple-comparison or Holm–Sidak post hoc analysis.
    Figure Legend Snippet: Expression of BI-1 improved long-term neurological deficits at 4 weeks post-HI. (A,B) In rat pups expressing BI-1 through treatment with Ad-TMBIM6 , long-term motor, learning and memory were assessed via foot fault test (A) and water maze test (B) at 4 weeks post-HI. (C,D) Quantitative analysis of percentage of weight of ipsilateral and contralateral regions out of total brain weights (C) and percentage of tissue loss (D) at 4 weeks post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; n =6–9. One-way ANOVA followed by Tukey multiple-comparison or Holm–Sidak post hoc analysis.

    Techniques Used: Expressing, Hi-C

    Effects of BI-1 siRNA and IRE1α CRISPR activation plasmid on infarct area at 72 h post-HI. (A,B) Representative images of infarcted tissue (A) and quantification of percentage infarcted area (out of total brain area) (B) at 72 h post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (C) Representative schematic of BI-1/IRE1α/XBP1/CHOP signaling pathway using the intervention groups (BI-1 siRNA and IRE1α CRISPR activation plasmid) to study the mechanism. (D) Representative immunofluorescence staining of BI-1 (red) with neurons (NeuN, green), astrocytes (GFAP, green) and microglia (Iba-1, green) after siRNA administration. Representative picture demonstrating brain region from where immunofluorescence pictures were obtained is shown upper right. n =1/group. Scale bars: 50 μm.
    Figure Legend Snippet: Effects of BI-1 siRNA and IRE1α CRISPR activation plasmid on infarct area at 72 h post-HI. (A,B) Representative images of infarcted tissue (A) and quantification of percentage infarcted area (out of total brain area) (B) at 72 h post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (C) Representative schematic of BI-1/IRE1α/XBP1/CHOP signaling pathway using the intervention groups (BI-1 siRNA and IRE1α CRISPR activation plasmid) to study the mechanism. (D) Representative immunofluorescence staining of BI-1 (red) with neurons (NeuN, green), astrocytes (GFAP, green) and microglia (Iba-1, green) after siRNA administration. Representative picture demonstrating brain region from where immunofluorescence pictures were obtained is shown upper right. n =1/group. Scale bars: 50 μm.

    Techniques Used: CRISPR, Activation Assay, Plasmid Preparation, Immunofluorescence, Staining

    BI-1 exerted its protective effects via the IRE1α–XBP1–CHOP pathway at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–F) Quantitative analysis of BI-1 (B), pIRE1α (C), XBP1 (D), CHOP (E) and ROMO1 (F) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR; ^ P <0.05 versus Ad-TMBIM6+BI-1 siRNA; ε P <0.05 versus BI-1 siRNA-only group. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.
    Figure Legend Snippet: BI-1 exerted its protective effects via the IRE1α–XBP1–CHOP pathway at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–F) Quantitative analysis of BI-1 (B), pIRE1α (C), XBP1 (D), CHOP (E) and ROMO1 (F) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR; ^ P <0.05 versus Ad-TMBIM6+BI-1 siRNA; ε P <0.05 versus BI-1 siRNA-only group. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Techniques Used: Western Blot, Expressing, CRISPR

    Overexpression of BI-1 attenuated apoptosis at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–D) Quantification of Bcl-2 (B), BAX (C) and CC3 (D) expression level data (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison, Holm–Sidak or Dunnett's post hoc analysis. (E) Representative microphotographs of positively CC3-stained (red) neurons (NeuN, green) from 10-day-old rats at 72 h post-HI. (F) Representative microphotographs of Fluoro-Jade C (FJC)-positive degenerating neurons in the peri-infarcted area. n =1/group. Scale bars: 100 µm.
    Figure Legend Snippet: Overexpression of BI-1 attenuated apoptosis at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–D) Quantification of Bcl-2 (B), BAX (C) and CC3 (D) expression level data (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison, Holm–Sidak or Dunnett's post hoc analysis. (E) Representative microphotographs of positively CC3-stained (red) neurons (NeuN, green) from 10-day-old rats at 72 h post-HI. (F) Representative microphotographs of Fluoro-Jade C (FJC)-positive degenerating neurons in the peri-infarcted area. n =1/group. Scale bars: 100 µm.

    Techniques Used: Over Expression, Western Blot, Expressing, CRISPR, Staining

    Percentage cell viability significantly improved after transfection of PC-12 cells with Ad-TMBIM6 at MOI=100 in an in vitro OGD model. (A) Quantification of data shows time-dependent decrease in percentage cell viability for cultured PC12 cells when exposed to 1 h, 1.5, 3 h, 5 h and 6 h in OGD. (B) Quantification analysis of percentage cell viability of PC12 cells when exposed to varying concentrations (MOI) of viral vector. (C) Representative images of cell morphology and density of cells after OGD in all groups. Scale bar: 100 µm. (D) Quantification analysis of percentage cell viability with pathway interventions. Data presented as mean± s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6+scramble siRNA (D) or versus MOI=100 (B); ε P <0.05 versus Ad-TMBIM6 only; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 versus DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.
    Figure Legend Snippet: Percentage cell viability significantly improved after transfection of PC-12 cells with Ad-TMBIM6 at MOI=100 in an in vitro OGD model. (A) Quantification of data shows time-dependent decrease in percentage cell viability for cultured PC12 cells when exposed to 1 h, 1.5, 3 h, 5 h and 6 h in OGD. (B) Quantification analysis of percentage cell viability of PC12 cells when exposed to varying concentrations (MOI) of viral vector. (C) Representative images of cell morphology and density of cells after OGD in all groups. Scale bar: 100 µm. (D) Quantification analysis of percentage cell viability with pathway interventions. Data presented as mean± s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6+scramble siRNA (D) or versus MOI=100 (B); ε P <0.05 versus Ad-TMBIM6 only; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 versus DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Techniques Used: Transfection, In Vitro, Cell Culture, Plasmid Preparation, CRISPR

    BI-1 exerted its anti-apoptotic effects via inhibition of IRE1α in an in vitro OGD model. (A) Representative immunoblots of protein expression levels in PC12 cultured cells. Quantification of BI-1 (B), pIRE1α (C), pPERK (D) and CC3 (E) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6 and Ad-TMBIM6+scramble siRNA; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.
    Figure Legend Snippet: BI-1 exerted its anti-apoptotic effects via inhibition of IRE1α in an in vitro OGD model. (A) Representative immunoblots of protein expression levels in PC12 cultured cells. Quantification of BI-1 (B), pIRE1α (C), pPERK (D) and CC3 (E) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6 and Ad-TMBIM6+scramble siRNA; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Techniques Used: Inhibition, In Vitro, Western Blot, Expressing, Cell Culture, CRISPR

    Related Articles

    CRISPR:

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model
    Article Snippet: Human adenoviral TMBIM6 vector (Ad-TMBIM6) (ADV-225645, Vector Biolabs) was injected intracerebroventricularly (icv) at 2 μl containing 1.6×10 11 PFU/ml per injection ( ) at 72 h, 48 h and 24 h pre-HI and 1 h post-HI. .. BI-1 siRNA (sc-270613, Santa Cruz Biotechnology) and IREα CRISPR activation plasmid (AP) (sc-400576-ACT, Santa Cruz Biotechnology) were administered icv at 48 h pre-HI. .. Scramble siRNA (sc-37007, Santa Cruz Biotechnology) and control CRISPR (sc-437275, Santa Cruz Biotechnology) were also administered to control animals.

    Activation Assay:

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model
    Article Snippet: Human adenoviral TMBIM6 vector (Ad-TMBIM6) (ADV-225645, Vector Biolabs) was injected intracerebroventricularly (icv) at 2 μl containing 1.6×10 11 PFU/ml per injection ( ) at 72 h, 48 h and 24 h pre-HI and 1 h post-HI. .. BI-1 siRNA (sc-270613, Santa Cruz Biotechnology) and IREα CRISPR activation plasmid (AP) (sc-400576-ACT, Santa Cruz Biotechnology) were administered icv at 48 h pre-HI. .. Scramble siRNA (sc-37007, Santa Cruz Biotechnology) and control CRISPR (sc-437275, Santa Cruz Biotechnology) were also administered to control animals.

    Plasmid Preparation:

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model
    Article Snippet: Human adenoviral TMBIM6 vector (Ad-TMBIM6) (ADV-225645, Vector Biolabs) was injected intracerebroventricularly (icv) at 2 μl containing 1.6×10 11 PFU/ml per injection ( ) at 72 h, 48 h and 24 h pre-HI and 1 h post-HI. .. BI-1 siRNA (sc-270613, Santa Cruz Biotechnology) and IREα CRISPR activation plasmid (AP) (sc-400576-ACT, Santa Cruz Biotechnology) were administered icv at 48 h pre-HI. .. Scramble siRNA (sc-37007, Santa Cruz Biotechnology) and control CRISPR (sc-437275, Santa Cruz Biotechnology) were also administered to control animals.



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    Image Search Results


    Expression levels of endogenous BI-1, IRE1α, XBP1 and CHOP post-HI. (A) Representative immunoblots showing protein expression levels in ipsilateral hemispheric tissue from 10-day-old neonatal rats. (B–E) Quantitative analysis of BI-1 (B), IRE1α (C), XBP1 (D) and CHOP (E) time-dependent expression after HI (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus 6 h, @ P <0.05 versus 24 h; n =4. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Expression levels of endogenous BI-1, IRE1α, XBP1 and CHOP post-HI. (A) Representative immunoblots showing protein expression levels in ipsilateral hemispheric tissue from 10-day-old neonatal rats. (B–E) Quantitative analysis of BI-1 (B), IRE1α (C), XBP1 (D) and CHOP (E) time-dependent expression after HI (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus 6 h, @ P <0.05 versus 24 h; n =4. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Article Snippet: Scramble siRNA (sc-37007, Santa Cruz Biotechnology) and siRNA targeting BI-1 (sc-270613, Santa Cruz Biotechnology) was prepared according to manufacturer's protocol.

    Techniques: Expressing, Western Blot

    Ad-TMBIM6 administered at 48 h pre-HI reduced percentage infarcted area and showed localization to neurons and microglia at 72 h post-HI. (A) Representative images of TTC-stained sections and quantification of percentage infarcted area at 72 h post-HI in brain tissue from neonatal rats expressing BI-1 through adenoviral transduction with Ad-TMBIM6. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle, n =6. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (B–E) Representative microphotographs of double immunofluorescence staining of BI-1 (red) with neurons (NeuN, green) (B), IRE1α (red) with neurons (NeuN, green) (C), BI-1 (red) with astrocytes (GFAP, green) (D) and BI-1 (red) with microglia (Iba-1, green) (E) in the peri-infarcted area at 72 h post-HI. Blue, DAPI. n =1/group. Merged images show localization of BI-1 or IRE1α to neurons, astrocytes or microglia. There was minimal localization of BI-1 to astrocytes. Scale bars: 50 μm.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Ad-TMBIM6 administered at 48 h pre-HI reduced percentage infarcted area and showed localization to neurons and microglia at 72 h post-HI. (A) Representative images of TTC-stained sections and quantification of percentage infarcted area at 72 h post-HI in brain tissue from neonatal rats expressing BI-1 through adenoviral transduction with Ad-TMBIM6. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle, n =6. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (B–E) Representative microphotographs of double immunofluorescence staining of BI-1 (red) with neurons (NeuN, green) (B), IRE1α (red) with neurons (NeuN, green) (C), BI-1 (red) with astrocytes (GFAP, green) (D) and BI-1 (red) with microglia (Iba-1, green) (E) in the peri-infarcted area at 72 h post-HI. Blue, DAPI. n =1/group. Merged images show localization of BI-1 or IRE1α to neurons, astrocytes or microglia. There was minimal localization of BI-1 to astrocytes. Scale bars: 50 μm.

    Article Snippet: Scramble siRNA (sc-37007, Santa Cruz Biotechnology) and siRNA targeting BI-1 (sc-270613, Santa Cruz Biotechnology) was prepared according to manufacturer's protocol.

    Techniques: Staining, Expressing, Transduction, Double Immunofluorescence Staining

    Expression of BI-1 improved long-term neurological deficits at 4 weeks post-HI. (A,B) In rat pups expressing BI-1 through treatment with Ad-TMBIM6 , long-term motor, learning and memory were assessed via foot fault test (A) and water maze test (B) at 4 weeks post-HI. (C,D) Quantitative analysis of percentage of weight of ipsilateral and contralateral regions out of total brain weights (C) and percentage of tissue loss (D) at 4 weeks post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; n =6–9. One-way ANOVA followed by Tukey multiple-comparison or Holm–Sidak post hoc analysis.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Expression of BI-1 improved long-term neurological deficits at 4 weeks post-HI. (A,B) In rat pups expressing BI-1 through treatment with Ad-TMBIM6 , long-term motor, learning and memory were assessed via foot fault test (A) and water maze test (B) at 4 weeks post-HI. (C,D) Quantitative analysis of percentage of weight of ipsilateral and contralateral regions out of total brain weights (C) and percentage of tissue loss (D) at 4 weeks post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; n =6–9. One-way ANOVA followed by Tukey multiple-comparison or Holm–Sidak post hoc analysis.

    Article Snippet: Scramble siRNA (sc-37007, Santa Cruz Biotechnology) and siRNA targeting BI-1 (sc-270613, Santa Cruz Biotechnology) was prepared according to manufacturer's protocol.

    Techniques: Expressing, Hi-C

    Effects of BI-1 siRNA and IRE1α CRISPR activation plasmid on infarct area at 72 h post-HI. (A,B) Representative images of infarcted tissue (A) and quantification of percentage infarcted area (out of total brain area) (B) at 72 h post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (C) Representative schematic of BI-1/IRE1α/XBP1/CHOP signaling pathway using the intervention groups (BI-1 siRNA and IRE1α CRISPR activation plasmid) to study the mechanism. (D) Representative immunofluorescence staining of BI-1 (red) with neurons (NeuN, green), astrocytes (GFAP, green) and microglia (Iba-1, green) after siRNA administration. Representative picture demonstrating brain region from where immunofluorescence pictures were obtained is shown upper right. n =1/group. Scale bars: 50 μm.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Effects of BI-1 siRNA and IRE1α CRISPR activation plasmid on infarct area at 72 h post-HI. (A,B) Representative images of infarcted tissue (A) and quantification of percentage infarcted area (out of total brain area) (B) at 72 h post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (C) Representative schematic of BI-1/IRE1α/XBP1/CHOP signaling pathway using the intervention groups (BI-1 siRNA and IRE1α CRISPR activation plasmid) to study the mechanism. (D) Representative immunofluorescence staining of BI-1 (red) with neurons (NeuN, green), astrocytes (GFAP, green) and microglia (Iba-1, green) after siRNA administration. Representative picture demonstrating brain region from where immunofluorescence pictures were obtained is shown upper right. n =1/group. Scale bars: 50 μm.

    Article Snippet: Scramble siRNA (sc-37007, Santa Cruz Biotechnology) and siRNA targeting BI-1 (sc-270613, Santa Cruz Biotechnology) was prepared according to manufacturer's protocol.

    Techniques: CRISPR, Activation Assay, Plasmid Preparation, Immunofluorescence, Staining

    BI-1 exerted its protective effects via the IRE1α–XBP1–CHOP pathway at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–F) Quantitative analysis of BI-1 (B), pIRE1α (C), XBP1 (D), CHOP (E) and ROMO1 (F) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR; ^ P <0.05 versus Ad-TMBIM6+BI-1 siRNA; ε P <0.05 versus BI-1 siRNA-only group. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: BI-1 exerted its protective effects via the IRE1α–XBP1–CHOP pathway at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–F) Quantitative analysis of BI-1 (B), pIRE1α (C), XBP1 (D), CHOP (E) and ROMO1 (F) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR; ^ P <0.05 versus Ad-TMBIM6+BI-1 siRNA; ε P <0.05 versus BI-1 siRNA-only group. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Article Snippet: Scramble siRNA (sc-37007, Santa Cruz Biotechnology) and siRNA targeting BI-1 (sc-270613, Santa Cruz Biotechnology) was prepared according to manufacturer's protocol.

    Techniques: Western Blot, Expressing, CRISPR

    Overexpression of BI-1 attenuated apoptosis at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–D) Quantification of Bcl-2 (B), BAX (C) and CC3 (D) expression level data (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison, Holm–Sidak or Dunnett's post hoc analysis. (E) Representative microphotographs of positively CC3-stained (red) neurons (NeuN, green) from 10-day-old rats at 72 h post-HI. (F) Representative microphotographs of Fluoro-Jade C (FJC)-positive degenerating neurons in the peri-infarcted area. n =1/group. Scale bars: 100 µm.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Overexpression of BI-1 attenuated apoptosis at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–D) Quantification of Bcl-2 (B), BAX (C) and CC3 (D) expression level data (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison, Holm–Sidak or Dunnett's post hoc analysis. (E) Representative microphotographs of positively CC3-stained (red) neurons (NeuN, green) from 10-day-old rats at 72 h post-HI. (F) Representative microphotographs of Fluoro-Jade C (FJC)-positive degenerating neurons in the peri-infarcted area. n =1/group. Scale bars: 100 µm.

    Article Snippet: Scramble siRNA (sc-37007, Santa Cruz Biotechnology) and siRNA targeting BI-1 (sc-270613, Santa Cruz Biotechnology) was prepared according to manufacturer's protocol.

    Techniques: Over Expression, Western Blot, Expressing, CRISPR, Staining

    Percentage cell viability significantly improved after transfection of PC-12 cells with Ad-TMBIM6 at MOI=100 in an in vitro OGD model. (A) Quantification of data shows time-dependent decrease in percentage cell viability for cultured PC12 cells when exposed to 1 h, 1.5, 3 h, 5 h and 6 h in OGD. (B) Quantification analysis of percentage cell viability of PC12 cells when exposed to varying concentrations (MOI) of viral vector. (C) Representative images of cell morphology and density of cells after OGD in all groups. Scale bar: 100 µm. (D) Quantification analysis of percentage cell viability with pathway interventions. Data presented as mean± s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6+scramble siRNA (D) or versus MOI=100 (B); ε P <0.05 versus Ad-TMBIM6 only; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 versus DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Percentage cell viability significantly improved after transfection of PC-12 cells with Ad-TMBIM6 at MOI=100 in an in vitro OGD model. (A) Quantification of data shows time-dependent decrease in percentage cell viability for cultured PC12 cells when exposed to 1 h, 1.5, 3 h, 5 h and 6 h in OGD. (B) Quantification analysis of percentage cell viability of PC12 cells when exposed to varying concentrations (MOI) of viral vector. (C) Representative images of cell morphology and density of cells after OGD in all groups. Scale bar: 100 µm. (D) Quantification analysis of percentage cell viability with pathway interventions. Data presented as mean± s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6+scramble siRNA (D) or versus MOI=100 (B); ε P <0.05 versus Ad-TMBIM6 only; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 versus DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Article Snippet: Scramble siRNA (sc-37007, Santa Cruz Biotechnology) and siRNA targeting BI-1 (sc-270613, Santa Cruz Biotechnology) was prepared according to manufacturer's protocol.

    Techniques: Transfection, In Vitro, Cell Culture, Plasmid Preparation, CRISPR

    BI-1 exerted its anti-apoptotic effects via inhibition of IRE1α in an in vitro OGD model. (A) Representative immunoblots of protein expression levels in PC12 cultured cells. Quantification of BI-1 (B), pIRE1α (C), pPERK (D) and CC3 (E) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6 and Ad-TMBIM6+scramble siRNA; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: BI-1 exerted its anti-apoptotic effects via inhibition of IRE1α in an in vitro OGD model. (A) Representative immunoblots of protein expression levels in PC12 cultured cells. Quantification of BI-1 (B), pIRE1α (C), pPERK (D) and CC3 (E) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6 and Ad-TMBIM6+scramble siRNA; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Article Snippet: Scramble siRNA (sc-37007, Santa Cruz Biotechnology) and siRNA targeting BI-1 (sc-270613, Santa Cruz Biotechnology) was prepared according to manufacturer's protocol.

    Techniques: Inhibition, In Vitro, Western Blot, Expressing, Cell Culture, CRISPR

    Expression levels of endogenous BI-1, IRE1α, XBP1 and CHOP post-HI. (A) Representative immunoblots showing protein expression levels in ipsilateral hemispheric tissue from 10-day-old neonatal rats. (B–E) Quantitative analysis of BI-1 (B), IRE1α (C), XBP1 (D) and CHOP (E) time-dependent expression after HI (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus 6 h, @ P <0.05 versus 24 h; n =4. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Expression levels of endogenous BI-1, IRE1α, XBP1 and CHOP post-HI. (A) Representative immunoblots showing protein expression levels in ipsilateral hemispheric tissue from 10-day-old neonatal rats. (B–E) Quantitative analysis of BI-1 (B), IRE1α (C), XBP1 (D) and CHOP (E) time-dependent expression after HI (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus 6 h, @ P <0.05 versus 24 h; n =4. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Article Snippet: BI-1 siRNA (sc-270613, Santa Cruz Biotechnology) and IREα CRISPR activation plasmid (AP) (sc-400576-ACT, Santa Cruz Biotechnology) were administered icv at 48 h pre-HI.

    Techniques: Expressing, Western Blot

    Ad-TMBIM6 administered at 48 h pre-HI reduced percentage infarcted area and showed localization to neurons and microglia at 72 h post-HI. (A) Representative images of TTC-stained sections and quantification of percentage infarcted area at 72 h post-HI in brain tissue from neonatal rats expressing BI-1 through adenoviral transduction with Ad-TMBIM6. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle, n =6. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (B–E) Representative microphotographs of double immunofluorescence staining of BI-1 (red) with neurons (NeuN, green) (B), IRE1α (red) with neurons (NeuN, green) (C), BI-1 (red) with astrocytes (GFAP, green) (D) and BI-1 (red) with microglia (Iba-1, green) (E) in the peri-infarcted area at 72 h post-HI. Blue, DAPI. n =1/group. Merged images show localization of BI-1 or IRE1α to neurons, astrocytes or microglia. There was minimal localization of BI-1 to astrocytes. Scale bars: 50 μm.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Ad-TMBIM6 administered at 48 h pre-HI reduced percentage infarcted area and showed localization to neurons and microglia at 72 h post-HI. (A) Representative images of TTC-stained sections and quantification of percentage infarcted area at 72 h post-HI in brain tissue from neonatal rats expressing BI-1 through adenoviral transduction with Ad-TMBIM6. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle, n =6. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (B–E) Representative microphotographs of double immunofluorescence staining of BI-1 (red) with neurons (NeuN, green) (B), IRE1α (red) with neurons (NeuN, green) (C), BI-1 (red) with astrocytes (GFAP, green) (D) and BI-1 (red) with microglia (Iba-1, green) (E) in the peri-infarcted area at 72 h post-HI. Blue, DAPI. n =1/group. Merged images show localization of BI-1 or IRE1α to neurons, astrocytes or microglia. There was minimal localization of BI-1 to astrocytes. Scale bars: 50 μm.

    Article Snippet: BI-1 siRNA (sc-270613, Santa Cruz Biotechnology) and IREα CRISPR activation plasmid (AP) (sc-400576-ACT, Santa Cruz Biotechnology) were administered icv at 48 h pre-HI.

    Techniques: Staining, Expressing, Transduction, Double Immunofluorescence Staining

    Expression of BI-1 improved long-term neurological deficits at 4 weeks post-HI. (A,B) In rat pups expressing BI-1 through treatment with Ad-TMBIM6 , long-term motor, learning and memory were assessed via foot fault test (A) and water maze test (B) at 4 weeks post-HI. (C,D) Quantitative analysis of percentage of weight of ipsilateral and contralateral regions out of total brain weights (C) and percentage of tissue loss (D) at 4 weeks post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; n =6–9. One-way ANOVA followed by Tukey multiple-comparison or Holm–Sidak post hoc analysis.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Expression of BI-1 improved long-term neurological deficits at 4 weeks post-HI. (A,B) In rat pups expressing BI-1 through treatment with Ad-TMBIM6 , long-term motor, learning and memory were assessed via foot fault test (A) and water maze test (B) at 4 weeks post-HI. (C,D) Quantitative analysis of percentage of weight of ipsilateral and contralateral regions out of total brain weights (C) and percentage of tissue loss (D) at 4 weeks post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; n =6–9. One-way ANOVA followed by Tukey multiple-comparison or Holm–Sidak post hoc analysis.

    Article Snippet: BI-1 siRNA (sc-270613, Santa Cruz Biotechnology) and IREα CRISPR activation plasmid (AP) (sc-400576-ACT, Santa Cruz Biotechnology) were administered icv at 48 h pre-HI.

    Techniques: Expressing, Hi-C

    Effects of BI-1 siRNA and IRE1α CRISPR activation plasmid on infarct area at 72 h post-HI. (A,B) Representative images of infarcted tissue (A) and quantification of percentage infarcted area (out of total brain area) (B) at 72 h post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (C) Representative schematic of BI-1/IRE1α/XBP1/CHOP signaling pathway using the intervention groups (BI-1 siRNA and IRE1α CRISPR activation plasmid) to study the mechanism. (D) Representative immunofluorescence staining of BI-1 (red) with neurons (NeuN, green), astrocytes (GFAP, green) and microglia (Iba-1, green) after siRNA administration. Representative picture demonstrating brain region from where immunofluorescence pictures were obtained is shown upper right. n =1/group. Scale bars: 50 μm.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Effects of BI-1 siRNA and IRE1α CRISPR activation plasmid on infarct area at 72 h post-HI. (A,B) Representative images of infarcted tissue (A) and quantification of percentage infarcted area (out of total brain area) (B) at 72 h post-HI. Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis. (C) Representative schematic of BI-1/IRE1α/XBP1/CHOP signaling pathway using the intervention groups (BI-1 siRNA and IRE1α CRISPR activation plasmid) to study the mechanism. (D) Representative immunofluorescence staining of BI-1 (red) with neurons (NeuN, green), astrocytes (GFAP, green) and microglia (Iba-1, green) after siRNA administration. Representative picture demonstrating brain region from where immunofluorescence pictures were obtained is shown upper right. n =1/group. Scale bars: 50 μm.

    Article Snippet: BI-1 siRNA (sc-270613, Santa Cruz Biotechnology) and IREα CRISPR activation plasmid (AP) (sc-400576-ACT, Santa Cruz Biotechnology) were administered icv at 48 h pre-HI.

    Techniques: CRISPR, Activation Assay, Plasmid Preparation, Immunofluorescence, Staining

    BI-1 exerted its protective effects via the IRE1α–XBP1–CHOP pathway at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–F) Quantitative analysis of BI-1 (B), pIRE1α (C), XBP1 (D), CHOP (E) and ROMO1 (F) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR; ^ P <0.05 versus Ad-TMBIM6+BI-1 siRNA; ε P <0.05 versus BI-1 siRNA-only group. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: BI-1 exerted its protective effects via the IRE1α–XBP1–CHOP pathway at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–F) Quantitative analysis of BI-1 (B), pIRE1α (C), XBP1 (D), CHOP (E) and ROMO1 (F) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR; ^ P <0.05 versus Ad-TMBIM6+BI-1 siRNA; ε P <0.05 versus BI-1 siRNA-only group. n =6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Article Snippet: BI-1 siRNA (sc-270613, Santa Cruz Biotechnology) and IREα CRISPR activation plasmid (AP) (sc-400576-ACT, Santa Cruz Biotechnology) were administered icv at 48 h pre-HI.

    Techniques: Western Blot, Expressing, CRISPR

    Overexpression of BI-1 attenuated apoptosis at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–D) Quantification of Bcl-2 (B), BAX (C) and CC3 (D) expression level data (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison, Holm–Sidak or Dunnett's post hoc analysis. (E) Representative microphotographs of positively CC3-stained (red) neurons (NeuN, green) from 10-day-old rats at 72 h post-HI. (F) Representative microphotographs of Fluoro-Jade C (FJC)-positive degenerating neurons in the peri-infarcted area. n =1/group. Scale bars: 100 µm.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Overexpression of BI-1 attenuated apoptosis at 72 h post-HI. (A) Representative immunoblots of protein expression levels in ipsilateral hemispheric tissue from 10-day-old rats at 72 h post-HI. (B–D) Quantification of Bcl-2 (B), BAX (C) and CC3 (D) expression level data (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus sham; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6; $ P <0.05 versus BI-1 siRNA; + P <0.05 versus Ad-TMBIM6+scramble siRNA; & P <0.05 versus IRE1α CRISPR AP; % P <0.05 versus control CRISPR. n =6/group. One-way ANOVA followed by Tukey multiple-comparison, Holm–Sidak or Dunnett's post hoc analysis. (E) Representative microphotographs of positively CC3-stained (red) neurons (NeuN, green) from 10-day-old rats at 72 h post-HI. (F) Representative microphotographs of Fluoro-Jade C (FJC)-positive degenerating neurons in the peri-infarcted area. n =1/group. Scale bars: 100 µm.

    Article Snippet: BI-1 siRNA (sc-270613, Santa Cruz Biotechnology) and IREα CRISPR activation plasmid (AP) (sc-400576-ACT, Santa Cruz Biotechnology) were administered icv at 48 h pre-HI.

    Techniques: Over Expression, Western Blot, Expressing, CRISPR, Staining

    Percentage cell viability significantly improved after transfection of PC-12 cells with Ad-TMBIM6 at MOI=100 in an in vitro OGD model. (A) Quantification of data shows time-dependent decrease in percentage cell viability for cultured PC12 cells when exposed to 1 h, 1.5, 3 h, 5 h and 6 h in OGD. (B) Quantification analysis of percentage cell viability of PC12 cells when exposed to varying concentrations (MOI) of viral vector. (C) Representative images of cell morphology and density of cells after OGD in all groups. Scale bar: 100 µm. (D) Quantification analysis of percentage cell viability with pathway interventions. Data presented as mean± s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6+scramble siRNA (D) or versus MOI=100 (B); ε P <0.05 versus Ad-TMBIM6 only; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 versus DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: Percentage cell viability significantly improved after transfection of PC-12 cells with Ad-TMBIM6 at MOI=100 in an in vitro OGD model. (A) Quantification of data shows time-dependent decrease in percentage cell viability for cultured PC12 cells when exposed to 1 h, 1.5, 3 h, 5 h and 6 h in OGD. (B) Quantification analysis of percentage cell viability of PC12 cells when exposed to varying concentrations (MOI) of viral vector. (C) Representative images of cell morphology and density of cells after OGD in all groups. Scale bar: 100 µm. (D) Quantification analysis of percentage cell viability with pathway interventions. Data presented as mean± s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6+scramble siRNA (D) or versus MOI=100 (B); ε P <0.05 versus Ad-TMBIM6 only; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 versus DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Article Snippet: BI-1 siRNA (sc-270613, Santa Cruz Biotechnology) and IREα CRISPR activation plasmid (AP) (sc-400576-ACT, Santa Cruz Biotechnology) were administered icv at 48 h pre-HI.

    Techniques: Transfection, In Vitro, Cell Culture, Plasmid Preparation, CRISPR

    BI-1 exerted its anti-apoptotic effects via inhibition of IRE1α in an in vitro OGD model. (A) Representative immunoblots of protein expression levels in PC12 cultured cells. Quantification of BI-1 (B), pIRE1α (C), pPERK (D) and CC3 (E) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6 and Ad-TMBIM6+scramble siRNA; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Journal: Disease Models & Mechanisms

    Article Title: Adenoviral TMBIM6 vector attenuates ER-stress-induced apoptosis in a neonatal hypoxic-ischemic rat model

    doi: 10.1242/dmm.040352

    Figure Lengend Snippet: BI-1 exerted its anti-apoptotic effects via inhibition of IRE1α in an in vitro OGD model. (A) Representative immunoblots of protein expression levels in PC12 cultured cells. Quantification of BI-1 (B), pIRE1α (C), pPERK (D) and CC3 (E) expression levels (band density relative to actin). Data presented as mean±s.d.; * P <0.05 versus control; # P <0.05 versus vehicle; @ P <0.05 versus Ad-TMBIM6 and Ad-TMBIM6+scramble siRNA; $ P <0.05 versus BI-1 siRNA; & P <0.05 versus APY-29; + P <0.05 DMSO; % P <0.05 versus CCT020312; ^ P <0.05 versus STF-083010 and control CRISPR. n =5–6/group. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis.

    Article Snippet: BI-1 siRNA (sc-270613, Santa Cruz Biotechnology) and IREα CRISPR activation plasmid (AP) (sc-400576-ACT, Santa Cruz Biotechnology) were administered icv at 48 h pre-HI.

    Techniques: Inhibition, In Vitro, Western Blot, Expressing, Cell Culture, CRISPR

    Endogenous expression profiles of BI-1, NPR, P4502E1, Nrf-2, and HO-1 post-HI. Representative western blot bands of the temporal expression of endogenous proteins in the ipsilateral hemisphere post-HI ( a ). Quantification data of western blot bands for BI-1 ( b ), NPR ( c ), P4502E1 ( d ), Nrf-2 ( e ), and HO-1 ( f ) (data expressed as mean ± SD; an asterisk indicates p < 0.05 vs sham; number sign p < 0.05 vs 6 h, @ p < 0.05 vs 24 h; n = 4, using one-way ANOVA followed by Tukey’s multiple-comparison post hoc analysis)

    Journal: Journal of Neuroinflammation

    Article Title: Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

    doi: 10.1186/s12974-019-1559-4

    Figure Lengend Snippet: Endogenous expression profiles of BI-1, NPR, P4502E1, Nrf-2, and HO-1 post-HI. Representative western blot bands of the temporal expression of endogenous proteins in the ipsilateral hemisphere post-HI ( a ). Quantification data of western blot bands for BI-1 ( b ), NPR ( c ), P4502E1 ( d ), Nrf-2 ( e ), and HO-1 ( f ) (data expressed as mean ± SD; an asterisk indicates p < 0.05 vs sham; number sign p < 0.05 vs 6 h, @ p < 0.05 vs 24 h; n = 4, using one-way ANOVA followed by Tukey’s multiple-comparison post hoc analysis)

    Article Snippet: Two microliters of BI-1 siRNA (300 pmol/μL, Sigma-Aldrich) or Nrf-2 siRNA or scramble siRNA (300 pmol/μL, Santa Cruz) were administered icv using a Hamilton syringe (10 μL, Hamilton Co) into the right lateral ventricle (1.5 mm posterior, 1.5 mm lateral to bregma and 1.7 mm down from the surface of the brain) at 48 h pre-HI at a rate of 0.3 μL/min [ ].

    Techniques: Expressing, Western Blot

    Silencing BI-1 or Nrf-2 increased percent infarcted area at 72 h Post-HI. Quantitative analysis of percent infarcted area after administration of BI-1 or Nrf-2 siRNA ( a , b ) (data expressed as mean ± SD; an asterisk indicates p < 0.05 vs sham; number sign p < 0.05 vs vehicle; @ p < 0.05 vs Ad-TMBIM6 or scramble; n = 6–8/group using one-way ANOVA followed by Tukey’s multiple-comparison post hoc analysis)

    Journal: Journal of Neuroinflammation

    Article Title: Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

    doi: 10.1186/s12974-019-1559-4

    Figure Lengend Snippet: Silencing BI-1 or Nrf-2 increased percent infarcted area at 72 h Post-HI. Quantitative analysis of percent infarcted area after administration of BI-1 or Nrf-2 siRNA ( a , b ) (data expressed as mean ± SD; an asterisk indicates p < 0.05 vs sham; number sign p < 0.05 vs vehicle; @ p < 0.05 vs Ad-TMBIM6 or scramble; n = 6–8/group using one-way ANOVA followed by Tukey’s multiple-comparison post hoc analysis)

    Article Snippet: Two microliters of BI-1 siRNA (300 pmol/μL, Sigma-Aldrich) or Nrf-2 siRNA or scramble siRNA (300 pmol/μL, Santa Cruz) were administered icv using a Hamilton syringe (10 μL, Hamilton Co) into the right lateral ventricle (1.5 mm posterior, 1.5 mm lateral to bregma and 1.7 mm down from the surface of the brain) at 48 h pre-HI at a rate of 0.3 μL/min [ ].

    Techniques:

    The effects of silencing BI-1 on the NPR-CYP complex at 72 h post-HI. Representative bands of western blot data ( a ). Quantification of western blot bands for BI-1 ( b ) and P4502E1 ( c ) (data expressed as mean ± SD; an asterisk indicates p < 0.05 vs sham; number sign p < 0.05 vs vehicle; @ p < 0.05 vs Ad-TMBIM6 or scramble; n = 6/group using one-way ANOVA followed by Tukey’s multiple-comparison post hoc analysis). Immunofluorescence staining of Iba-1 with BI-1 or P4502E1 ( d , e ). Green was for microglial staining, red was for BI-1 or P4502E1 staining, and blue was for DAPI. Merge (yellow) showed the co-localization of BI-1 or P4502E1 on microglia (scale bar = 50 μm; n = 3/group)

    Journal: Journal of Neuroinflammation

    Article Title: Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

    doi: 10.1186/s12974-019-1559-4

    Figure Lengend Snippet: The effects of silencing BI-1 on the NPR-CYP complex at 72 h post-HI. Representative bands of western blot data ( a ). Quantification of western blot bands for BI-1 ( b ) and P4502E1 ( c ) (data expressed as mean ± SD; an asterisk indicates p < 0.05 vs sham; number sign p < 0.05 vs vehicle; @ p < 0.05 vs Ad-TMBIM6 or scramble; n = 6/group using one-way ANOVA followed by Tukey’s multiple-comparison post hoc analysis). Immunofluorescence staining of Iba-1 with BI-1 or P4502E1 ( d , e ). Green was for microglial staining, red was for BI-1 or P4502E1 staining, and blue was for DAPI. Merge (yellow) showed the co-localization of BI-1 or P4502E1 on microglia (scale bar = 50 μm; n = 3/group)

    Article Snippet: Two microliters of BI-1 siRNA (300 pmol/μL, Sigma-Aldrich) or Nrf-2 siRNA or scramble siRNA (300 pmol/μL, Santa Cruz) were administered icv using a Hamilton syringe (10 μL, Hamilton Co) into the right lateral ventricle (1.5 mm posterior, 1.5 mm lateral to bregma and 1.7 mm down from the surface of the brain) at 48 h pre-HI at a rate of 0.3 μL/min [ ].

    Techniques: Western Blot, Immunofluorescence, Staining

    Overexpression of BI-1 attenuated ROS production and inflammation at 72 h post-HI. Representative picture of western blot bands ( a ). Quantification analysis of band intensities for ROMO1 ( b ), IL-6 ( c ), TNFα ( d ), and IL-1β ( e ). Immunofluorescent staining and quantification of the number of positively stained cells of Iba-1 with the inflammatory marker, IL-1β, in the peri-infarcted region ( f ). Green was for microglial staining and red was for IL-1β. Merge showed the co-localization of IL-1β on microglia. Scale bar 50 μm; n = 3/group. Data expressed as mean ± SD; an asterisk indicates p < 0.05 vs sham; number sign p < 0.05 vs vehicle; @ p < 0.05 vs Ad-TMBIM6 or scramble; n = 6/group using one-way ANOVA followed by Tukey’s multiple-comparison post hoc analysis

    Journal: Journal of Neuroinflammation

    Article Title: Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

    doi: 10.1186/s12974-019-1559-4

    Figure Lengend Snippet: Overexpression of BI-1 attenuated ROS production and inflammation at 72 h post-HI. Representative picture of western blot bands ( a ). Quantification analysis of band intensities for ROMO1 ( b ), IL-6 ( c ), TNFα ( d ), and IL-1β ( e ). Immunofluorescent staining and quantification of the number of positively stained cells of Iba-1 with the inflammatory marker, IL-1β, in the peri-infarcted region ( f ). Green was for microglial staining and red was for IL-1β. Merge showed the co-localization of IL-1β on microglia. Scale bar 50 μm; n = 3/group. Data expressed as mean ± SD; an asterisk indicates p < 0.05 vs sham; number sign p < 0.05 vs vehicle; @ p < 0.05 vs Ad-TMBIM6 or scramble; n = 6/group using one-way ANOVA followed by Tukey’s multiple-comparison post hoc analysis

    Article Snippet: Two microliters of BI-1 siRNA (300 pmol/μL, Sigma-Aldrich) or Nrf-2 siRNA or scramble siRNA (300 pmol/μL, Santa Cruz) were administered icv using a Hamilton syringe (10 μL, Hamilton Co) into the right lateral ventricle (1.5 mm posterior, 1.5 mm lateral to bregma and 1.7 mm down from the surface of the brain) at 48 h pre-HI at a rate of 0.3 μL/min [ ].

    Techniques: Over Expression, Western Blot, Staining, Marker

    BI-1 exerts its anti-inflammatory effects via inhibition of P4502E1 in an in vitro OGD model. Representative picture of western blot bands ( a ). Quantification analysis of BI-1, IL-6, TNFα, IL-1β, P4502E1, pNrf-2 ( b – g ) and percent cell viability ( h ). Data represent ± SD; an asterisk indicates p < 0.05 vs control; number sign p < 0.05 vs vehicle; @ p < 0.05 vs Ad-TMBIM6 and scramble; dollar sign p < 0.05 vs BI-1 siRNA; n = 4–5/group using one-way ANOVA followed by Tukey’s multiple-comparison post hoc analysis

    Journal: Journal of Neuroinflammation

    Article Title: Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

    doi: 10.1186/s12974-019-1559-4

    Figure Lengend Snippet: BI-1 exerts its anti-inflammatory effects via inhibition of P4502E1 in an in vitro OGD model. Representative picture of western blot bands ( a ). Quantification analysis of BI-1, IL-6, TNFα, IL-1β, P4502E1, pNrf-2 ( b – g ) and percent cell viability ( h ). Data represent ± SD; an asterisk indicates p < 0.05 vs control; number sign p < 0.05 vs vehicle; @ p < 0.05 vs Ad-TMBIM6 and scramble; dollar sign p < 0.05 vs BI-1 siRNA; n = 4–5/group using one-way ANOVA followed by Tukey’s multiple-comparison post hoc analysis

    Article Snippet: Two microliters of BI-1 siRNA (300 pmol/μL, Sigma-Aldrich) or Nrf-2 siRNA or scramble siRNA (300 pmol/μL, Santa Cruz) were administered icv using a Hamilton syringe (10 μL, Hamilton Co) into the right lateral ventricle (1.5 mm posterior, 1.5 mm lateral to bregma and 1.7 mm down from the surface of the brain) at 48 h pre-HI at a rate of 0.3 μL/min [ ].

    Techniques: Inhibition, In Vitro, Western Blot