xbp1 Search Results


gene exp xbp1 mm00457357 m1  (Thermo Fisher)
96
Thermo Fisher gene exp xbp1 mm00457357 m1
(a) Spliced <t>Xbp1</t> mRNA normalized to total Xbp1 mRNA (sXbp1/Xbp1) in BMDCs treated with LPS, L-LA or MitoPQ for 6h. (b) sXPB1/XBP1 in human DCs treated with LPS, D-LA or L-LA for 6h. (c-e) XBP1 biding sites and XBP1 recruitment to Il1b (c), Il6 (d) and Il23 (e) promoters in BMDCs determined by ChIP after treatment with LPS, D-LA or L-LA for 6h. (f) sXbp1/Xbp1 in WT and HIF-1α Itgax splenic DCs at EAE peak. (g) sXbp1/Xbp1 in BMDCs over-expressing Ndufa4l2 . (h) EAE in WT (n=15) and XBP1 Itgax (n=15) mice. Experiment repeated three times. (i) IFN-γ + , IFN-γ + IL-17 + , and IL-17 + CD4 + T cells in CNS. (j,k) Recall cytokine (j) and proliferative (k) response following stimulation of WT and Xbp1 Itgax splenocytes with 20 μg/mL of MOG35-55 for 72h. n=5 mice per group. (l,m) IPA (l) and heatmap (m) of RNA-seq analysis of WT and Xbp1 Itgax splenic DCs 30 days after EAE induction. n=5 mice per group. Data shown as mean±SEM. ***p<0.001, **p<0.01, *p<0.05, ns: p>0.05.
Gene Exp Xbp1 Mm00457357 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gene exp xbp1 mm00457357 m1/product/Thermo Fisher
Average 96 stars, based on 1 article reviews
gene exp xbp1 mm00457357 m1 - by Bioz Stars, 2026-03
96/100 stars
  Buy from Supplier
xbp1 novus biologicals nbp177681ss  (Novus Biologicals)
93
Novus Biologicals xbp1 novus biologicals nbp177681ss
(a) Spliced <t>Xbp1</t> mRNA normalized to total Xbp1 mRNA (sXbp1/Xbp1) in BMDCs treated with LPS, L-LA or MitoPQ for 6h. (b) sXPB1/XBP1 in human DCs treated with LPS, D-LA or L-LA for 6h. (c-e) XBP1 biding sites and XBP1 recruitment to Il1b (c), Il6 (d) and Il23 (e) promoters in BMDCs determined by ChIP after treatment with LPS, D-LA or L-LA for 6h. (f) sXbp1/Xbp1 in WT and HIF-1α Itgax splenic DCs at EAE peak. (g) sXbp1/Xbp1 in BMDCs over-expressing Ndufa4l2 . (h) EAE in WT (n=15) and XBP1 Itgax (n=15) mice. Experiment repeated three times. (i) IFN-γ + , IFN-γ + IL-17 + , and IL-17 + CD4 + T cells in CNS. (j,k) Recall cytokine (j) and proliferative (k) response following stimulation of WT and Xbp1 Itgax splenocytes with 20 μg/mL of MOG35-55 for 72h. n=5 mice per group. (l,m) IPA (l) and heatmap (m) of RNA-seq analysis of WT and Xbp1 Itgax splenic DCs 30 days after EAE induction. n=5 mice per group. Data shown as mean±SEM. ***p<0.001, **p<0.01, *p<0.05, ns: p>0.05.
Xbp1 Novus Biologicals Nbp177681ss, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/xbp1 novus biologicals nbp177681ss/product/Novus Biologicals
Average 93 stars, based on 1 article reviews
xbp1 novus biologicals nbp177681ss - by Bioz Stars, 2026-03
93/100 stars
  Buy from Supplier
xbp1 mneongreen  (Addgene inc)
93
Addgene inc xbp1 mneongreen
Figure 3. Flow cytometric determination of the level of <t>XBP1</t> <t>(mNeonGreen)</t> protein in heat-stressed U2OS cells. Cells were heat-stressed at 40 ◦C or 42 ◦C for one hour, followed by six hours of recovery at 37 ◦C. The Kolmogorov–Smirnov test was performed to analyze the equality of distributions. Samples treated at 40 or 42 ◦C differed from the control (37 ◦C) significantly (p < 0.05).
Xbp1 Mneongreen, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/xbp1 mneongreen/product/Addgene inc
Average 93 stars, based on 1 article reviews
xbp1 mneongreen - by Bioz Stars, 2026-03
93/100 stars
  Buy from Supplier
x box binding protein 1  (Proteintech)
95
Proteintech x box binding protein 1
Figure 3. Flow cytometric determination of the level of <t>XBP1</t> <t>(mNeonGreen)</t> protein in heat-stressed U2OS cells. Cells were heat-stressed at 40 ◦C or 42 ◦C for one hour, followed by six hours of recovery at 37 ◦C. The Kolmogorov–Smirnov test was performed to analyze the equality of distributions. Samples treated at 40 or 42 ◦C differed from the control (37 ◦C) significantly (p < 0.05).
X Box Binding Protein 1, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/x box binding protein 1/product/Proteintech
Average 95 stars, based on 1 article reviews
x box binding protein 1 - by Bioz Stars, 2026-03
95/100 stars
  Buy from Supplier
anti xbp 1  (Santa Cruz Biotechnology)
95
Santa Cruz Biotechnology anti xbp 1
Figure 3. Flow cytometric determination of the level of <t>XBP1</t> <t>(mNeonGreen)</t> protein in heat-stressed U2OS cells. Cells were heat-stressed at 40 ◦C or 42 ◦C for one hour, followed by six hours of recovery at 37 ◦C. The Kolmogorov–Smirnov test was performed to analyze the equality of distributions. Samples treated at 40 or 42 ◦C differed from the control (37 ◦C) significantly (p < 0.05).
Anti Xbp 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti xbp 1/product/Santa Cruz Biotechnology
Average 95 stars, based on 1 article reviews
anti xbp 1 - by Bioz Stars, 2026-03
95/100 stars
  Buy from Supplier
gene exp xbp1 hs03929085 g1  (Thermo Fisher)
96
Thermo Fisher gene exp xbp1 hs03929085 g1
Figure 3. Flow cytometric determination of the level of <t>XBP1</t> <t>(mNeonGreen)</t> protein in heat-stressed U2OS cells. Cells were heat-stressed at 40 ◦C or 42 ◦C for one hour, followed by six hours of recovery at 37 ◦C. The Kolmogorov–Smirnov test was performed to analyze the equality of distributions. Samples treated at 40 or 42 ◦C differed from the control (37 ◦C) significantly (p < 0.05).
Gene Exp Xbp1 Hs03929085 G1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gene exp xbp1 hs03929085 g1/product/Thermo Fisher
Average 96 stars, based on 1 article reviews
gene exp xbp1 hs03929085 g1 - by Bioz Stars, 2026-03
96/100 stars
  Buy from Supplier
anti xbp1s  (Proteintech)
95
Proteintech anti xbp1s
Figure 3. Flow cytometric determination of the level of <t>XBP1</t> <t>(mNeonGreen)</t> protein in heat-stressed U2OS cells. Cells were heat-stressed at 40 ◦C or 42 ◦C for one hour, followed by six hours of recovery at 37 ◦C. The Kolmogorov–Smirnov test was performed to analyze the equality of distributions. Samples treated at 40 or 42 ◦C differed from the control (37 ◦C) significantly (p < 0.05).
Anti Xbp1s, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti xbp1s/product/Proteintech
Average 95 stars, based on 1 article reviews
anti xbp1s - by Bioz Stars, 2026-03
95/100 stars
  Buy from Supplier
rabbit polyclonal anti xbp1  (OriGene)
91
OriGene rabbit polyclonal anti xbp1
Figure 3. Flow cytometric determination of the level of <t>XBP1</t> <t>(mNeonGreen)</t> protein in heat-stressed U2OS cells. Cells were heat-stressed at 40 ◦C or 42 ◦C for one hour, followed by six hours of recovery at 37 ◦C. The Kolmogorov–Smirnov test was performed to analyze the equality of distributions. Samples treated at 40 or 42 ◦C differed from the control (37 ◦C) significantly (p < 0.05).
Rabbit Polyclonal Anti Xbp1, supplied by OriGene, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit polyclonal anti xbp1/product/OriGene
Average 91 stars, based on 1 article reviews
rabbit polyclonal anti xbp1 - by Bioz Stars, 2026-03
91/100 stars
  Buy from Supplier
gene exp xbp1 hs00231936 m1  (Thermo Fisher)
95
Thermo Fisher gene exp xbp1 hs00231936 m1
Figure 3. Flow cytometric determination of the level of <t>XBP1</t> <t>(mNeonGreen)</t> protein in heat-stressed U2OS cells. Cells were heat-stressed at 40 ◦C or 42 ◦C for one hour, followed by six hours of recovery at 37 ◦C. The Kolmogorov–Smirnov test was performed to analyze the equality of distributions. Samples treated at 40 or 42 ◦C differed from the control (37 ◦C) significantly (p < 0.05).
Gene Exp Xbp1 Hs00231936 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gene exp xbp1 hs00231936 m1/product/Thermo Fisher
Average 95 stars, based on 1 article reviews
gene exp xbp1 hs00231936 m1 - by Bioz Stars, 2026-03
95/100 stars
  Buy from Supplier
gene exp xbp1 mm03464496 m1  (Thermo Fisher)
92
Thermo Fisher gene exp xbp1 mm03464496 m1
(A and B) Heatmap of the mRNAs (A, blue/red) and proteins (B, blue/yellow) involved in the UPR pathway are shown. Proteomic and RNA sequencing data were obtained from EpCAM+ sorted epithelial cells from control and Emc3-cKO mice at E18.5. Genes and proteins were categorized by ToppGene. P values and fold changes for each mRNA and protein are listed in Supplemental Table 2. (C–H) Immunohistochemical staining for ATF3 (C and D), ATF4 (E and F), and ATF6 (G and H) was performed on lung sections from E18.5 control and Emc3-cKO embryos. ATF3 and ATF4 staining was increased and ATF6 staining was unaltered in the mutant lungs. Scale bars: 100 μm. (I) Western blots using EpCAM+ cell lysates from control and mutant lungs at E18.5 were performed using the indicated antibodies. (J) Increased <t>Xbp1</t> splicing in Emc3-cKO mice. Levels of the spliced Xbp1 transcript [Xbp1(S)] were normalized to that of the full-length Xbp1 by qPCR. mRNAs were isolated from E18.5 control and Emc3-cKO EpCAM+ cells. Data are the mean ± SEM. *P < 0.05 using unpaired, 2-tailed Student’s t test. n = 4/group. (K) Model for the induction of UPR in Emc3-cKO AT2 cells. The model was built based on the integration of RNA sequencing and proteomic data. Relationships between differentially expressed genes and proteins were determined by Genomatix Pathway System (GePS) and Ingenuity Pathway Analysis (IPA) suites. System models were created using IPA’s Path Designer.
Gene Exp Xbp1 Mm03464496 M1, supplied by Thermo Fisher, 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/result/gene exp xbp1 mm03464496 m1/product/Thermo Fisher
Average 92 stars, based on 1 article reviews
gene exp xbp1 mm03464496 m1 - by Bioz Stars, 2026-03
92/100 stars
  Buy from Supplier

Image Search Results


(a) Spliced Xbp1 mRNA normalized to total Xbp1 mRNA (sXbp1/Xbp1) in BMDCs treated with LPS, L-LA or MitoPQ for 6h. (b) sXPB1/XBP1 in human DCs treated with LPS, D-LA or L-LA for 6h. (c-e) XBP1 biding sites and XBP1 recruitment to Il1b (c), Il6 (d) and Il23 (e) promoters in BMDCs determined by ChIP after treatment with LPS, D-LA or L-LA for 6h. (f) sXbp1/Xbp1 in WT and HIF-1α Itgax splenic DCs at EAE peak. (g) sXbp1/Xbp1 in BMDCs over-expressing Ndufa4l2 . (h) EAE in WT (n=15) and XBP1 Itgax (n=15) mice. Experiment repeated three times. (i) IFN-γ + , IFN-γ + IL-17 + , and IL-17 + CD4 + T cells in CNS. (j,k) Recall cytokine (j) and proliferative (k) response following stimulation of WT and Xbp1 Itgax splenocytes with 20 μg/mL of MOG35-55 for 72h. n=5 mice per group. (l,m) IPA (l) and heatmap (m) of RNA-seq analysis of WT and Xbp1 Itgax splenic DCs 30 days after EAE induction. n=5 mice per group. Data shown as mean±SEM. ***p<0.001, **p<0.01, *p<0.05, ns: p>0.05.

Journal: bioRxiv

Article Title: Engineered probiotics limit CNS autoimmunity by stabilizing HIF-1α in dendritic cells

doi: 10.1101/2023.03.17.532101

Figure Lengend Snippet: (a) Spliced Xbp1 mRNA normalized to total Xbp1 mRNA (sXbp1/Xbp1) in BMDCs treated with LPS, L-LA or MitoPQ for 6h. (b) sXPB1/XBP1 in human DCs treated with LPS, D-LA or L-LA for 6h. (c-e) XBP1 biding sites and XBP1 recruitment to Il1b (c), Il6 (d) and Il23 (e) promoters in BMDCs determined by ChIP after treatment with LPS, D-LA or L-LA for 6h. (f) sXbp1/Xbp1 in WT and HIF-1α Itgax splenic DCs at EAE peak. (g) sXbp1/Xbp1 in BMDCs over-expressing Ndufa4l2 . (h) EAE in WT (n=15) and XBP1 Itgax (n=15) mice. Experiment repeated three times. (i) IFN-γ + , IFN-γ + IL-17 + , and IL-17 + CD4 + T cells in CNS. (j,k) Recall cytokine (j) and proliferative (k) response following stimulation of WT and Xbp1 Itgax splenocytes with 20 μg/mL of MOG35-55 for 72h. n=5 mice per group. (l,m) IPA (l) and heatmap (m) of RNA-seq analysis of WT and Xbp1 Itgax splenic DCs 30 days after EAE induction. n=5 mice per group. Data shown as mean±SEM. ***p<0.001, **p<0.01, *p<0.05, ns: p>0.05.

Article Snippet: Taqman probes used in this study are: Gapdh (Mm99999915_g1), Il1b (Mm00434228_m1), Il6 (Mm00446190_m1), Il12 (Mm00434169_m1), Il23 (Mm00518984_m1), Tnf (Mm00443528_m1), Ndufa4l2 (Mm01160374_g1), sXbp1 (Mm03464496_m1), Xbp1 (Mm00457357_m1), Gclm (Mm01324400_m1), Gls (Mm01257297_m1), Phgdh (Mm01623589_g1), Shmt2 (Mm00659512_g1), Slc7a11 (Mm00442530_m1), Lonp1 (Mm01236887_m1), Cox4i1 (Mm0250094_m1), Cox4i2 (Mm00446387_m1), Hif1a (Mm00468869_m1), Cd36 (Mm00432403_m1), Vhl (Mm00494137_m1), Mct1 (Mm01306379_m1), Slc2a1 (Mm00441480_m1), GAPDH (Hs02786624_g1), IL23 (Hs00372324_m1), IL1B (Hs01555410_m1), IL6 (Hs00174131_m1), HIF1a (Hs00153153_m1), IFNG (Hs00989291_m1), CSF2 (Hs00929873_m1). qPCR data were analyzed by the ddCt method by normalizing the expression of each gene to Gapdh and then to the control group.

Techniques: Expressing, RNA Sequencing

(a,b) sXbp1/Xbp1 in BMDCs treated with LPS or LPS+D-LA (a) and LPS, MitoPQ or MitoTempo (MitoTP) (b) for 6h. (c,d) XBP1 recruitment to Il1b , Il6 and Il23 promoters in BMDCs treated with LPS and mitoPQ (c) or LPS and ML228 (d) for 6h. (e) IFNγ + and IL-17 + CD4 splenic T cells in WT and Xbp1 Itgax mice 30 days after EAE induction. (f,g) Heatmap (f) and IPA (g) in CNS DCs from WT and Xbp1 Itgax mice. Data shown as mean±SEM. ***p<0.001, **p<0.01, *p<0.05, ns: p>0.05.

Journal: bioRxiv

Article Title: Engineered probiotics limit CNS autoimmunity by stabilizing HIF-1α in dendritic cells

doi: 10.1101/2023.03.17.532101

Figure Lengend Snippet: (a,b) sXbp1/Xbp1 in BMDCs treated with LPS or LPS+D-LA (a) and LPS, MitoPQ or MitoTempo (MitoTP) (b) for 6h. (c,d) XBP1 recruitment to Il1b , Il6 and Il23 promoters in BMDCs treated with LPS and mitoPQ (c) or LPS and ML228 (d) for 6h. (e) IFNγ + and IL-17 + CD4 splenic T cells in WT and Xbp1 Itgax mice 30 days after EAE induction. (f,g) Heatmap (f) and IPA (g) in CNS DCs from WT and Xbp1 Itgax mice. Data shown as mean±SEM. ***p<0.001, **p<0.01, *p<0.05, ns: p>0.05.

Article Snippet: Taqman probes used in this study are: Gapdh (Mm99999915_g1), Il1b (Mm00434228_m1), Il6 (Mm00446190_m1), Il12 (Mm00434169_m1), Il23 (Mm00518984_m1), Tnf (Mm00443528_m1), Ndufa4l2 (Mm01160374_g1), sXbp1 (Mm03464496_m1), Xbp1 (Mm00457357_m1), Gclm (Mm01324400_m1), Gls (Mm01257297_m1), Phgdh (Mm01623589_g1), Shmt2 (Mm00659512_g1), Slc7a11 (Mm00442530_m1), Lonp1 (Mm01236887_m1), Cox4i1 (Mm0250094_m1), Cox4i2 (Mm00446387_m1), Hif1a (Mm00468869_m1), Cd36 (Mm00432403_m1), Vhl (Mm00494137_m1), Mct1 (Mm01306379_m1), Slc2a1 (Mm00441480_m1), GAPDH (Hs02786624_g1), IL23 (Hs00372324_m1), IL1B (Hs01555410_m1), IL6 (Hs00174131_m1), HIF1a (Hs00153153_m1), IFNG (Hs00989291_m1), CSF2 (Hs00929873_m1). qPCR data were analyzed by the ddCt method by normalizing the expression of each gene to Gapdh and then to the control group.

Techniques:

(a) Engineered EcN Lac strain. (b) D-LA and L-LA in culture supernatants following EcN Lac or EcN incubation at 37°C. (c) D-LA in small intestine tissue after EcN Lac administration. (d,e) HIF-1α, NDUFA4L2 and spliced XBP1 protein staining (d) and quantification (e) in CD11c + cells from small intestine after EcN Lac or EcN gavage. (f) EAE development and (g) IFN-γ + , IL-17 + and IFN-γ + IL-17 + CD4 + T cell quantification in CNS of WT and HIF-1α Itgax mice treated with EcN or EcN Lac . Experiment repeated three times. (h) IFN-γ + and IL-17 + CD4 + T cell quantification in small intestine of WT mice treated with EcN or EcN Lac . ( i,j) Il1b, Il6, Il23, Tnf (i) and sXbp1/Xbp1 (j) expression in small intestine DCs from EcN or EcN Lac treated mice. (k) Representative dot plot of Kaede photoconversion in T cells from spleen. (l) Total, IFN-γ + and IL-17 + CD4 + T cell photoconverted in spleen of WT mice treated with EcN or EcN Lac . Data shown as mean±SEM. **p<0.01, *p<0.05, ns: p>0.05.

Journal: bioRxiv

Article Title: Engineered probiotics limit CNS autoimmunity by stabilizing HIF-1α in dendritic cells

doi: 10.1101/2023.03.17.532101

Figure Lengend Snippet: (a) Engineered EcN Lac strain. (b) D-LA and L-LA in culture supernatants following EcN Lac or EcN incubation at 37°C. (c) D-LA in small intestine tissue after EcN Lac administration. (d,e) HIF-1α, NDUFA4L2 and spliced XBP1 protein staining (d) and quantification (e) in CD11c + cells from small intestine after EcN Lac or EcN gavage. (f) EAE development and (g) IFN-γ + , IL-17 + and IFN-γ + IL-17 + CD4 + T cell quantification in CNS of WT and HIF-1α Itgax mice treated with EcN or EcN Lac . Experiment repeated three times. (h) IFN-γ + and IL-17 + CD4 + T cell quantification in small intestine of WT mice treated with EcN or EcN Lac . ( i,j) Il1b, Il6, Il23, Tnf (i) and sXbp1/Xbp1 (j) expression in small intestine DCs from EcN or EcN Lac treated mice. (k) Representative dot plot of Kaede photoconversion in T cells from spleen. (l) Total, IFN-γ + and IL-17 + CD4 + T cell photoconverted in spleen of WT mice treated with EcN or EcN Lac . Data shown as mean±SEM. **p<0.01, *p<0.05, ns: p>0.05.

Article Snippet: Taqman probes used in this study are: Gapdh (Mm99999915_g1), Il1b (Mm00434228_m1), Il6 (Mm00446190_m1), Il12 (Mm00434169_m1), Il23 (Mm00518984_m1), Tnf (Mm00443528_m1), Ndufa4l2 (Mm01160374_g1), sXbp1 (Mm03464496_m1), Xbp1 (Mm00457357_m1), Gclm (Mm01324400_m1), Gls (Mm01257297_m1), Phgdh (Mm01623589_g1), Shmt2 (Mm00659512_g1), Slc7a11 (Mm00442530_m1), Lonp1 (Mm01236887_m1), Cox4i1 (Mm0250094_m1), Cox4i2 (Mm00446387_m1), Hif1a (Mm00468869_m1), Cd36 (Mm00432403_m1), Vhl (Mm00494137_m1), Mct1 (Mm01306379_m1), Slc2a1 (Mm00441480_m1), GAPDH (Hs02786624_g1), IL23 (Hs00372324_m1), IL1B (Hs01555410_m1), IL6 (Hs00174131_m1), HIF1a (Hs00153153_m1), IFNG (Hs00989291_m1), CSF2 (Hs00929873_m1). qPCR data were analyzed by the ddCt method by normalizing the expression of each gene to Gapdh and then to the control group.

Techniques: Incubation, Staining, Expressing

Figure 3. Flow cytometric determination of the level of XBP1 (mNeonGreen) protein in heat-stressed U2OS cells. Cells were heat-stressed at 40 ◦C or 42 ◦C for one hour, followed by six hours of recovery at 37 ◦C. The Kolmogorov–Smirnov test was performed to analyze the equality of distributions. Samples treated at 40 or 42 ◦C differed from the control (37 ◦C) significantly (p < 0.05).

Journal: Cells

Article Title: Mild Hyperthermia-Induced Thermogenesis in the Endoplasmic Reticulum Defines Stress Response Mechanisms.

doi: 10.3390/cells13131141

Figure Lengend Snippet: Figure 3. Flow cytometric determination of the level of XBP1 (mNeonGreen) protein in heat-stressed U2OS cells. Cells were heat-stressed at 40 ◦C or 42 ◦C for one hour, followed by six hours of recovery at 37 ◦C. The Kolmogorov–Smirnov test was performed to analyze the equality of distributions. Samples treated at 40 or 42 ◦C differed from the control (37 ◦C) significantly (p < 0.05).

Article Snippet: To construct the U2OS cell line expressing fluorescently labeled XBP1 (mNeonGreen), 200 ng linearized pLHCX-XBP1 mNeonGreen plasmid NLS (Addgene plasmid #115971; http://n2t.net/addgene:115971; RRID: Addgene_115971), a gift from David Andrews [23], was transfected into 106 U2OS cells using the Amaxa Nucleofector System with the Cell Line Nucleofector Kit V (catalog no: VCA-1003, Lonza, Cologne, Germany) utilizing program X-001 according to the manufacturer’s protocol.

Techniques: Control

(A and B) Heatmap of the mRNAs (A, blue/red) and proteins (B, blue/yellow) involved in the UPR pathway are shown. Proteomic and RNA sequencing data were obtained from EpCAM+ sorted epithelial cells from control and Emc3-cKO mice at E18.5. Genes and proteins were categorized by ToppGene. P values and fold changes for each mRNA and protein are listed in Supplemental Table 2. (C–H) Immunohistochemical staining for ATF3 (C and D), ATF4 (E and F), and ATF6 (G and H) was performed on lung sections from E18.5 control and Emc3-cKO embryos. ATF3 and ATF4 staining was increased and ATF6 staining was unaltered in the mutant lungs. Scale bars: 100 μm. (I) Western blots using EpCAM+ cell lysates from control and mutant lungs at E18.5 were performed using the indicated antibodies. (J) Increased Xbp1 splicing in Emc3-cKO mice. Levels of the spliced Xbp1 transcript [Xbp1(S)] were normalized to that of the full-length Xbp1 by qPCR. mRNAs were isolated from E18.5 control and Emc3-cKO EpCAM+ cells. Data are the mean ± SEM. *P < 0.05 using unpaired, 2-tailed Student’s t test. n = 4/group. (K) Model for the induction of UPR in Emc3-cKO AT2 cells. The model was built based on the integration of RNA sequencing and proteomic data. Relationships between differentially expressed genes and proteins were determined by Genomatix Pathway System (GePS) and Ingenuity Pathway Analysis (IPA) suites. System models were created using IPA’s Path Designer.

Journal: The Journal of Clinical Investigation

Article Title: EMC3 coordinates surfactant protein and lipid homeostasis required for respiration

doi: 10.1172/JCI94152

Figure Lengend Snippet: (A and B) Heatmap of the mRNAs (A, blue/red) and proteins (B, blue/yellow) involved in the UPR pathway are shown. Proteomic and RNA sequencing data were obtained from EpCAM+ sorted epithelial cells from control and Emc3-cKO mice at E18.5. Genes and proteins were categorized by ToppGene. P values and fold changes for each mRNA and protein are listed in Supplemental Table 2. (C–H) Immunohistochemical staining for ATF3 (C and D), ATF4 (E and F), and ATF6 (G and H) was performed on lung sections from E18.5 control and Emc3-cKO embryos. ATF3 and ATF4 staining was increased and ATF6 staining was unaltered in the mutant lungs. Scale bars: 100 μm. (I) Western blots using EpCAM+ cell lysates from control and mutant lungs at E18.5 were performed using the indicated antibodies. (J) Increased Xbp1 splicing in Emc3-cKO mice. Levels of the spliced Xbp1 transcript [Xbp1(S)] were normalized to that of the full-length Xbp1 by qPCR. mRNAs were isolated from E18.5 control and Emc3-cKO EpCAM+ cells. Data are the mean ± SEM. *P < 0.05 using unpaired, 2-tailed Student’s t test. n = 4/group. (K) Model for the induction of UPR in Emc3-cKO AT2 cells. The model was built based on the integration of RNA sequencing and proteomic data. Relationships between differentially expressed genes and proteins were determined by Genomatix Pathway System (GePS) and Ingenuity Pathway Analysis (IPA) suites. System models were created using IPA’s Path Designer.

Article Snippet: The levels of the spliced form of XBP1 (TapMan probe Mm03464496_m1; Applied Biosystems) was normalized to that of the unspliced XBP1 (TapMan probe Mm03464497_s1; Applied Biosystems).

Techniques: RNA Sequencing, Control, Immunohistochemical staining, Staining, Mutagenesis, Western Blot, Isolation