rabbit polyclonal anti xbp1  (Novus Biologicals)

Journal: iScience

Article Title: Cetylpyridinium chloride triggers paraptosis to suppress pancreatic tumor growth via the ERN1-MAP3K5-p38 pathway

doi: 10.1016/j.isci.2024.110598

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Article Snippet: Rabbit polyclonal anti-XBP1s , Proteintech Biotechnology , Cat# 24868-1-AP; RRID: AB_2879766.

Techniques: Recombinant, Lysis, Modification, Polyacrylamide Gel Electrophoresis, Protease Inhibitor, CCK-8 Assay, Sequencing, Software

Journal: Frontiers in Physiology

Article Title: Constitutive expression of spliced X-box binding protein 1 inhibits dentin formation in mice

doi: 10.3389/fphys.2023.1319954

Figure Lengend Snippet: IRE1α activation-independent expression of XBP1S. (A, B) . Shown are the schematic representations of the Xbp1 minigene (A) and Xbp1s minigene (B) constructs. The Xbp1s minigene contains a modified exon 4 (Δ26) that lack the 26 intronic sequence, and it also contains a loxP site in intron 3. +1, the transcription start site; E, exon; green boxes, coding exons; and grey boxes, non-coding exons. AscI, NsiI, EcoRV, HindIII, SspI and AflII are restriction endonucleases. (C, D) . HEK 293 cells were transiently transfected with the Xbp1 minigene (C) or Xbp1s minigene (D) together with the pCDNA3 empty vector (Vector) (lane 3) or a construct expressing normal (Norm) IRE1α (lane 4) or two IRE1α variants, K599A (lane 5) and N906A (lane 6). HEK 293 cells were transfected with the pCDNA3 empty vector alone (lane 1) or the construct expressing normal IRE1α alone (lane 2) as controls. Total cell lysate was harvested 48 h after transfection and analyzed by Western-blotting with an antibody that recognizes both XBP1U and XBP1S, the blot was then stripped and sequentially probed with an antibody against phosphorylated IRE1α and total IRE1α. The blot was probed with mouse monoclonal β-actin antibody as the loading control.

Article Snippet: The membrane was sequentially immunoblotted with rabbit anti-XBP1 polyclonal antibody that recognizes both XBP1U and XBP1S (1:4000, Abcam, Cambridge, MA), horseradish peroxidase (HRP)-conjugated rabbit polyclonal anti-phosphorylated IRE1α (pSer724) (1:5000; Novus Biologicals) and mouse monoclonal anti-IRE1α antibody (1:1000; Santa Cruz Biotechnology, Inc.).

Techniques: Activation Assay, Expressing, Construct, Modification, Sequencing, Transfection, Plasmid Preparation, Western Blot

Journal: Frontiers in Physiology

Article Title: Constitutive expression of spliced X-box binding protein 1 inhibits dentin formation in mice

doi: 10.3389/fphys.2023.1319954

Figure Lengend Snippet: Immunohistochemical staining of total XBP1 (XBP1U and XBP1S) and XBP1S. Shown are the representative images of IHC staining of total XBP1 (including XBP1U and XBP1S) (A) ; signal in brown) and XBP1S (B) ; signal in brown) in the mandibular first molars of 3-week-old Xbp1 CS/+ and Twist2-Cre;Xbp1 CS/+ mice. Each image in (A, B) is from the middle region of the crown of a sagittally-sectioned mandibular first molar. (A1-A3, B1-B3) are the higher magnification views of the roof-forming odontoblasts (box1), central dental pulp cells (box 2) and floor-forming odontoblasts (box 3) in (A, B) , respectively. rd, roof dentin; fd, floor dentin; rod, roof-forming odontoblasts; fod, floor-forming odontoblasts. Note that the signals for total XBP1 were found in the dental pulps of Xbp1 CS/+ and Twist2-Cre;Xbp1 CS/+ mice, whereas the signals for XBP1S were strongly detected in the dental pulps of Twist2-Cre;Xbp1 CS/+ mice, but were barely detectable in those of Xbp1 CS/+ mice. Scale bars: 50 μm in (A, B) ; 20 μm in (A1-A3, B1-B3) .

Article Snippet: The membrane was sequentially immunoblotted with rabbit anti-XBP1 polyclonal antibody that recognizes both XBP1U and XBP1S (1:4000, Abcam, Cambridge, MA), horseradish peroxidase (HRP)-conjugated rabbit polyclonal anti-phosphorylated IRE1α (pSer724) (1:5000; Novus Biologicals) and mouse monoclonal anti-IRE1α antibody (1:1000; Santa Cruz Biotechnology, Inc.).

Techniques: Immunohistochemistry, Staining

Journal: Frontiers in Physiology

Article Title: Constitutive expression of spliced X-box binding protein 1 inhibits dentin formation in mice

doi: 10.3389/fphys.2023.1319954

Figure Lengend Snippet: P lain X-ray radiography and micro-computed tomography (µCT) analyses of the mandibular molars. (A, B) . Representative plain X-ray radiographs of the mandibular molars of 3-week-old and 7-week-old Xbp1 CS/+ and Twist2-Cre;Xbp1 CS/+ mice. M1, first molar; M2, second molar; M3, third molar. Scale bar: 500 μm. (C, D) . Representative 3-dimensional reconstructed μCT images (sagittal sections) of the mandibular first molars of 3-week-old and 7-week-old Xbp1 CS/+ and Twist2-Cre;Xbp1 CS/+ mice. The mesial side of each molar is on the right, and the distal side is on the left. Scale bar: 200 μm.

Article Snippet: The membrane was sequentially immunoblotted with rabbit anti-XBP1 polyclonal antibody that recognizes both XBP1U and XBP1S (1:4000, Abcam, Cambridge, MA), horseradish peroxidase (HRP)-conjugated rabbit polyclonal anti-phosphorylated IRE1α (pSer724) (1:5000; Novus Biologicals) and mouse monoclonal anti-IRE1α antibody (1:1000; Santa Cruz Biotechnology, Inc.).

Techniques: Micro-CT

Journal: Frontiers in Physiology

Article Title: Constitutive expression of spliced X-box binding protein 1 inhibits dentin formation in mice

doi: 10.3389/fphys.2023.1319954

Figure Lengend Snippet: Quantitative µCT analysis of mandibular first molars. µCT analysis was performed to quantify the roof dentin thickness (A) , floor dentin thickness (B) , pulp volume (C) , dentin/cementum volume (D) and dentin/cementum density (E) of mandibular first molars of 3- and 7-week-old mice. Student’s t test was used to compare the difference between Xbp1 CS/+ and Twist2-Cre;Xbp1 CS/+ mice. All values are mean ± SD. n = 3 for each group in A-B; n = 5 for each group in C-E; * p < 0.05; ** p < 0.01.

Article Snippet: The membrane was sequentially immunoblotted with rabbit anti-XBP1 polyclonal antibody that recognizes both XBP1U and XBP1S (1:4000, Abcam, Cambridge, MA), horseradish peroxidase (HRP)-conjugated rabbit polyclonal anti-phosphorylated IRE1α (pSer724) (1:5000; Novus Biologicals) and mouse monoclonal anti-IRE1α antibody (1:1000; Santa Cruz Biotechnology, Inc.).

Techniques:

Journal: Frontiers in Physiology

Article Title: Constitutive expression of spliced X-box binding protein 1 inhibits dentin formation in mice

doi: 10.3389/fphys.2023.1319954

Figure Lengend Snippet: H&E staining of the mandibular first molars. (A) . Shown are the representative images of H&E staining of a sagittally-sectioned mandibular first molars of 3-week-old Xbp1 CS/+ and Twist2-Cre;Xbp1 CS/+ mice. (A1, A2) are the higher magnification views of the roof-forming odontoblasts (box1) and floor-forming odontoblasts (box 2) in A. Note that the roof-forming odontoblasts were long, columnar-shaped and highly polarized in Xbp1 CS/+ mice, but the roof-forming odontoblasts became shorter and irregular in Twist2-Cre;Xbp1 CS/+ mice. ree, reduced enamel epithelium; pd, predentin, rd, roof dentin; fd, floor dentin; rod, roof-forming odontoblasts; fod, floor-forming odontoblasts. Scale bars: 200 μm in A; 20 μm in (A1, A2) .

Article Snippet: The membrane was sequentially immunoblotted with rabbit anti-XBP1 polyclonal antibody that recognizes both XBP1U and XBP1S (1:4000, Abcam, Cambridge, MA), horseradish peroxidase (HRP)-conjugated rabbit polyclonal anti-phosphorylated IRE1α (pSer724) (1:5000; Novus Biologicals) and mouse monoclonal anti-IRE1α antibody (1:1000; Santa Cruz Biotechnology, Inc.).

Techniques: Staining

Journal: Frontiers in Physiology

Article Title: Constitutive expression of spliced X-box binding protein 1 inhibits dentin formation in mice

doi: 10.3389/fphys.2023.1319954

Figure Lengend Snippet: In situ hybridization analyses of DSPP and DMP1 mRNA. Shown are the representative in situ hybridization analyses of DSPP mRNA (A) ; signal in purple) and DMP1 mRNA (B) ; signal in purple) in the mandibular first molars of 3-week-old Xbp1 CS/+ and Twist2-Cre;Xbp1 CS/+ mice. Each image in (A, B) is from the middle region of the crown of a sagittally-sectioned mandibular first molar. (A1-A2, B1-B2) are the higher magnification views of the roof-forming odontoblasts (box1) and floor-forming odontoblasts (box 2) in (A, B) , respectively. rod, roof-forming odontoblasts; fod, floor-forming odontoblasts. Scale bars: 200 μm in ( A, B) ; 20 μm in (A1-A2, B1-B2) .

Article Snippet: The membrane was sequentially immunoblotted with rabbit anti-XBP1 polyclonal antibody that recognizes both XBP1U and XBP1S (1:4000, Abcam, Cambridge, MA), horseradish peroxidase (HRP)-conjugated rabbit polyclonal anti-phosphorylated IRE1α (pSer724) (1:5000; Novus Biologicals) and mouse monoclonal anti-IRE1α antibody (1:1000; Santa Cruz Biotechnology, Inc.).

Techniques: In Situ Hybridization

Journal: Frontiers in Physiology

Article Title: Constitutive expression of spliced X-box binding protein 1 inhibits dentin formation in mice

doi: 10.3389/fphys.2023.1319954

Figure Lengend Snippet: Immunohistochemical staining of DSP/DSPP and DMP1 protein. Shown are the representative images of IHC staining of DSP/DSPP (A) ; signal in brown) and DMP1 (B) ; signal in brown) in the mandibular first molars of 3-week-old Xbp1 CS/+ and Twist2-Cre;Xbp1 CS/+ mice. Each image in (A, B) is from the middle region of the crown of a sagittally-sectioned mandibular first molar. (A1-A2, B1-B2) are the higher magnification views of box 1 and box 2 in (A, B) , respectively. rd, roof dentin; fd, floor dentin; rod, roof-forming odontoblasts; fod, floor-forming odontoblasts. Scale bars: 200 μm in (A, B) ; 20 μm in (A1-A2, B1-B2) .

Article Snippet: The membrane was sequentially immunoblotted with rabbit anti-XBP1 polyclonal antibody that recognizes both XBP1U and XBP1S (1:4000, Abcam, Cambridge, MA), horseradish peroxidase (HRP)-conjugated rabbit polyclonal anti-phosphorylated IRE1α (pSer724) (1:5000; Novus Biologicals) and mouse monoclonal anti-IRE1α antibody (1:1000; Santa Cruz Biotechnology, Inc.).

Techniques: Immunohistochemistry, Staining