Journal: Heliyon

Article Title: PSO/SDF-1 composite hydrogel promotes osteogenic differentiation of PDLSCs and bone regeneration in periodontitis rats.

doi: 10.1016/j.heliyon.2024.e32686

Figure Lengend Snippet: Fig. 6. Expression of osteogenic genes of PDLSCs by the hydrogels. (A) The mRNA expression level of ALP, RUNX2 and OCN. (B) The protein expression level of ALP, RUNX2, and OCN. Data are expressed as mean ± SD, N = 3. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 vs Control group.

Article Snippet: Rabbit anti-ALP antibody (1:100, 18176-1-AP, Proteintech), RUNX2 (1:100, ab236639, Abcam), and OCN (1:100, 23418-1-AP, Proteintech) were used to incubated on sections overnight at 4 ◦C.

Techniques: Expressing, Control

Journal: Heliyon

Article Title: PSO/SDF-1 composite hydrogel promotes osteogenic differentiation of PDLSCs and bone regeneration in periodontitis rats.

doi: 10.1016/j.heliyon.2024.e32686

Figure Lengend Snippet: Fig. 8. IHC analysis of different hydrogels. (A) Representative pictures of IHC of ALP. The scale bar is 500 μm and 100 μm. (B) Representative pictures of IHC of RUNX2. The scale bar is 500 μm and 100 μm. (C) Representative pictures of IHC of OCN. The scale bar is 500 μm and 100 μm. (D) IHC quantitative analysis of ALP. (E) IHC quantitative analysis of RUNX2. (F) IHC quantitative analysis of OCN. Data are expressed as mean ± SD, N = 6. ###, P < 0.001; ####, P < 0.0001 vs Model group.

Article Snippet: Rabbit anti-ALP antibody (1:100, 18176-1-AP, Proteintech), RUNX2 (1:100, ab236639, Abcam), and OCN (1:100, 23418-1-AP, Proteintech) were used to incubated on sections overnight at 4 ◦C.

Techniques:

Journal:

Article Title: Aryl hydrocarbon (Ah) receptor levels are selectively modulated by hsp90-associated immunophilin homolog XAP2

doi:

Figure Lengend Snippet: XAP2 is expressed in various established cell lines. Eight different cell lines were cultured, harvested, and homogenized. Total cytosolic extracts were isolated, and 75 μg of each lysate was resolved by SDS-PAGE, transferred to PVDF membrane, and analyzed by immunoblot analysis. XAP2 was detected with an XAP2 polyclonal antibody as primary antibody and [125I]-DAR as secondary antibody

Article Snippet: Rabbit polyclonal anti-XAP2 antibodies were provided by E. Croze (Berlex Laboratories).

Techniques: Cell Culture, Isolation, SDS Page, Membrane, Western Blot

Journal:

Article Title: Aryl hydrocarbon (Ah) receptor levels are selectively modulated by hsp90-associated immunophilin homolog XAP2

doi:

Figure Lengend Snippet: XAP2 specifically enhances the level of AhR in COS-1 cells when compared to other hsp90-binding TPR-containing proteins. COS-1 cells were transfected in 6 well dishes with 1 μg of pcDNA3/βmAhR and 0, 0.2, 0.4, 0.6, 0.8, or 1.0 μg of pCI/XAP2, pCI/FKBP52, pCMV6/PP5-FLAG, or pCMV6/PP5-TPR-FLAG (shaded triangle) and brought to a total of 2 μg vector/dish with pCI vector. (A) Total cell lysates were isolated, and 75 μg of each lysate were resolved by SDS-PAGE, transferred to PVDF membrane, and analyzed by immunoblot analysis. XAP2 and FKBP52 were detected with XAP2 polyclonal and FKBP52 monoclonal antibodies. PP5-TPR and PP5-TPR-FLAG were detected with anti-FLAG M2 antibody. AhR was detected with the anti-AhR monoclonal antibody RPT1. XAP2 was visualized with [125I]-DAR, and the AhR, FKBP52, PP5-FLAG, and PP5-TPR-FLAG were visualized with [125I]-SAM. (B) The graph depicts the fold change in AhR levels obtained in the presence of TPR-containing proteins after phosphorimaging of the blots. This experiment has been repeated 3 times with essentially the same results

Article Snippet: Rabbit polyclonal anti-XAP2 antibodies were provided by E. Croze (Berlex Laboratories).

Techniques: Binding Assay, Transfection, Plasmid Preparation, Isolation, SDS Page, Membrane, Western Blot, Bioprocessing

Journal:

Article Title: Aryl hydrocarbon (Ah) receptor levels are selectively modulated by hsp90-associated immunophilin homolog XAP2

doi:

Figure Lengend Snippet: Schematic representation of XAP2 TPR mutants and their ability to bind to hsp90 in COS1 cells. (A) Top panel: Black boxes indicate amino acid substitutions in the respective mutant. Bottom panel: Black boxes indicate conserved amino acid residues between XAP2 and FKBP52. The TPR consensus was generated from CDC16, CDC23, CDC27, SSN6, and SK13 (Lamb et al 1995). (B) COS-1 cells were transiently transfected with pCI/XAP2-FLAG, pCI/XAP2-FLAG-TPR mutants, or pCI (control), and cell lysate was isolated, immunoabsorbed with the M2 anti-FLAG affinity resin, eluted with FLAG peptide, resolved by SDS-PAGE, and transferred to PVDF membrane, followed by immunoblot analysis. Hsp90 was visualized with polyclonal antibodies raised against hsp84/86 and [125I]-DAR, and XAP2-FLAG was visualized with anti-FLAG M2 antibody and [125I]-SAM

Article Snippet: Rabbit polyclonal anti-XAP2 antibodies were provided by E. Croze (Berlex Laboratories).

Techniques: Mutagenesis, Generated, Transfection, Control, Isolation, SDS Page, Membrane, Western Blot

Journal:

Article Title: Aryl hydrocarbon (Ah) receptor levels are selectively modulated by hsp90-associated immunophilin homolog XAP2

doi:

Figure Lengend Snippet: Mutants G272D, G272E, and F288A are unable to interact with the AhR in the absence of hsp90 in a cell-free system, and a conserved glycine in the XAP2 TPR complex is required for assembly of XAP2 in AhR-hsp90 complexes in COS-1 cells. (A) XAP2-FLAG, XAP2-FLAG-TPR mutants, or control (RL) were labeled with [35S] methionine in RL independently, immunoabsorbed with M2 resin, and eluted with FLAG peptide. mAhR was generated in RL (unlabeled) and immunoabsorbed with RPT9/protein G sepharose or with murine IgG/protein G sepharose (control, last 2 lanes). The AhR immunoabsorption was washed in PBS to remove hsp90. RPT9-sepharose-mAhR was mixed with eluted XAP2-FLAG or XAP2-FLAG TPR mutants, and complexes washed, resolved by SDS-PAGE, and transferred to PVDF membrane. Top panel: mAhR visualized with RPT1 and [125I]-SAM by autoradiography. Middle panel: XAP2-FLAG visualized by autoradiography. (B) COS-1 cells were transiently cotransfected with pCI/XAP2-FLAG, pCI/XAP2-FLAG-TPR mutants, or pCI (control) and pcDNA3/_mAhR, and cell lysate was isolated, immunoabsorbed with the M2 anti-FLAG affinity resin, eluted with FLAG peptide, resolved by SDS-PAGE, and transferred to PVDF membrane, followed by immunoblot analysis. AhR was visualized with RPT1 and [125I]-SAM, hsp90 was visualized with polyclonal antibodies raised against hsp84/86 and [125I]-DAR, and XAP2-FLAG was visualized with anti-FLAG M2 antibody and [125I]-SAM.><<002>><<002>><<002>>>

Article Snippet: Rabbit polyclonal anti-XAP2 antibodies were provided by E. Croze (Berlex Laboratories).

Techniques: Control, Labeling, Generated, SDS Page, Membrane, Autoradiography, Isolation, Western Blot

Journal:

Article Title: Aryl hydrocarbon (Ah) receptor levels are selectively modulated by hsp90-associated immunophilin homolog XAP2

doi:

Figure Lengend Snippet: XAP2 mutants G272D and G272E are defective in enhancing the level of AhR in COS-1 cells. COS-1 cells were transfected in 6 well dishes with 1 μg of pcDNA3/βmAhR and 0, 0.2, 0.4, 0.6, 0.8, or 1.0 μg pCI/XAP2-G272D or pCI/XAP2-G272E or with 0, 0.25, 0.5, or 1.0 μg pCI/XAP2. The vector pCI was used to bring each transfection to a total of 2 μg plasmid/dish. Total cell lysates were isolated, and 75 μg of each lysate were resolved by SDS-PAGE, transferred to PVDF membrane, and analyzed by immunoblot analysis. XAP2 was detected with XAP2 polyclonal antibodies, and AhR was detected with the anti-AhR monoclonal antibody RPT1. XAP2 antibody was visualized with [125I]-DAR, and AhR antibody was visualized with [125I]-SAM. The graph depicts the fold change in AhR levels obtained in the presence of TPR-containing proteins after phosphorimaging of the blots. This experiment was performed twice with essentially the same results

Article Snippet: Rabbit polyclonal anti-XAP2 antibodies were provided by E. Croze (Berlex Laboratories).

Techniques: Transfection, Plasmid Preparation, Isolation, SDS Page, Membrane, Western Blot

Journal:

Article Title: Aryl hydrocarbon (Ah) receptor levels are selectively modulated by hsp90-associated immunophilin homolog XAP2

doi:

Figure Lengend Snippet: Cellular XAP2 is unaffected by geldanamycin or radicicol treatment. Hepa-1 cells were treated with GA or radicicol at 100 nM and 2 μM for 6 h. Following treatment, cytosol was isolated, and 100 μg were resolved by SDS-PAGE and transferred to PVDF membrane, followed by immunoblot analysis with antibodies specific for the AhR, XAP2, and FKBP52. AhR and FKBP52 were visualized with [125I]-SAM and XAP2 with [125I]-DAR and subsequent autoradiography

Article Snippet: Rabbit polyclonal anti-XAP2 antibodies were provided by E. Croze (Berlex Laboratories).

Techniques: Isolation, SDS Page, Membrane, Western Blot, Autoradiography

Journal:

Article Title: Aryl hydrocarbon (Ah) receptor levels are selectively modulated by hsp90-associated immunophilin homolog XAP2

doi:

Figure Lengend Snippet: EYFP-XAP2 binds to hsp90 and to AhR/hsp90 complexes in COS-1 cells. Upper panels: pEYFP-XAP2-FLAG or pEYFP alone was transiently transfected in COS-1 cells, cytosol was isolated, and 150 μg of protein were resolved by SDS-PAGE and transferred to PVDF, followed by immunoblot analysis with XAP2 polyclonal antibodies or anti-GFP monoclonal antibodies. Antibodies were visualized with DAR-P and GAM-P, respectively. Lower panels: pEYFP-XAP2-FLAG or pEYFP were transiently transfected in COS-1 cells (left panel); pEYFP-XAP2-FLAG or pEYFP were transiently cotransfected with pcDNA3/βmAhR in COS-1 cells (right panel). In both experiments, cytosol was isolated and immunoabsorbed with the M2 affinity resin, and complexes were eluted with FLAG peptide and resolved by SDS-PAGE, transferred to PVDF membrane, and analyzed by immunoblot analysis. pEYFP-XAP2-FLAG was visualized with the M2 antibody, AhR with the RPT1 antibody, and hsp90 with rabbit polyclonal antibodies against hsp84 and hsp86. M2 antibody was visualized with GAM-P, hsp90 with DAR-P, and AhR with GAM-P by ECL

Article Snippet: Rabbit polyclonal anti-XAP2 antibodies were provided by E. Croze (Berlex Laboratories).

Techniques: Transfection, Isolation, SDS Page, Western Blot, Bioprocessing, Membrane

Journal:

Article Title: Aryl hydrocarbon (Ah) receptor levels are selectively modulated by hsp90-associated immunophilin homolog XAP2

doi:

Figure Lengend Snippet: YFP-XAP2/Flag and YFP-XAP2/Flag G272D are localized in both the cytoplasmic and nuclear compartments. NIH 3T3 cells were transiently transfected with pEYFP (top panels), pEYFP-XAP2/Flag (middle panels), or pEYFP-XAP2 G272D (bottom panels). After 24 h, the transfected cells were visualized by either epifluorecence (left panels) or scanning confocal microscopy (right panels). The confocal images represent a horizontal midsectional view of the cell. Each image is representative of the population of transfected cells

Article Snippet: Rabbit polyclonal anti-XAP2 antibodies were provided by E. Croze (Berlex Laboratories).

Techniques: Transfection, Confocal Microscopy