b 27 Search Results


smc 165  (StressMarq)
90
StressMarq smc 165
Smc 165, supplied by StressMarq, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/smc 165/product/StressMarq
Average 90 stars, based on 1 article reviews
smc 165 - by Bioz Stars, 2026-04
90/100 stars
  Buy from Supplier
amphotericin b  (Chem Impex International)
95
Chem Impex International amphotericin b
Amphotericin B, supplied by Chem Impex International, 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/amphotericin b/product/Chem Impex International
Average 95 stars, based on 1 article reviews
amphotericin b - by Bioz Stars, 2026-04
95/100 stars
  Buy from Supplier
hspb5  (StressMarq)
90
StressMarq hspb5
Canonical members of HSPB family prevent the aggregation of Aβ-GFP. Cells were transfected with HSPB1, <t>HSPB5,</t> HSPB7 or FRTTO at 1:3 ratio for 48 h. Quantification of the pellet/soluble ratio of Aβ-GFP relative to FRTTO was depicted in the chart above the blot. Values represent mean ± SE of two independent experiments. ** P < 0.01, ns = non significant.
Hspb5, supplied by StressMarq, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/hspb5/product/StressMarq
Average 90 stars, based on 1 article reviews
hspb5 - by Bioz Stars, 2026-04
90/100 stars
  Buy from Supplier
s 3 amino 3 4 hydroxyphenyl propanoic acid  (Chem Impex International)
95
Chem Impex International s 3 amino 3 4 hydroxyphenyl propanoic acid
Canonical members of HSPB family prevent the aggregation of Aβ-GFP. Cells were transfected with HSPB1, <t>HSPB5,</t> HSPB7 or FRTTO at 1:3 ratio for 48 h. Quantification of the pellet/soluble ratio of Aβ-GFP relative to FRTTO was depicted in the chart above the blot. Values represent mean ± SE of two independent experiments. ** P < 0.01, ns = non significant.
S 3 Amino 3 4 Hydroxyphenyl Propanoic Acid, supplied by Chem Impex International, 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/s 3 amino 3 4 hydroxyphenyl propanoic acid/product/Chem Impex International
Average 95 stars, based on 1 article reviews
s 3 amino 3 4 hydroxyphenyl propanoic acid - by Bioz Stars, 2026-04
95/100 stars
  Buy from Supplier
trimethoprim  (Chem Impex International)
95
Chem Impex International trimethoprim
(A) Thiomaltose <t>trimethoprim</t> (TM-TMP): A new trimethoprim prodrug with enhanced water solubility that targets bacteria. (B) Proposed cleavage of TM-TMP in the presence of exogenous thiols.
Trimethoprim, supplied by Chem Impex International, 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/trimethoprim/product/Chem Impex International
Average 95 stars, based on 1 article reviews
trimethoprim - by Bioz Stars, 2026-04
95/100 stars
  Buy from Supplier
anti hla b27  (OriGene)
90
OriGene anti hla b27
(A) ARS staining of mineralization in AS MSCs transduced with shHLA-B or shCtrl under osteogenic induction with quantification (B). (C) Immunoblot analyses showing the expression of RARB and TNAP in AS MSCs expressing shHLA-B or shCtrl at day 7 under osteogenic induction. (D) Immunoblot showing <t>HLA-B27</t> expressions of control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. (E) ARS staining of control MSCs transduced with control lentiviral vector (pLAS2w) or a vector expressing HLA-B27 (pLAS2w-HLA-B27) with quantification (F). (G) Immunoblot analyses showing RARB and TNAP expressions in control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. All experiments done in the AS patient group and the control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or 1-way ANOVA, followed by Tukey’s HSD test. Representative images from AS (A1) MSCs are shown in C. Scale bars: 200 μm (A and E).
Anti Hla B27, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti hla b27/product/OriGene
Average 90 stars, based on 1 article reviews
anti hla b27 - by Bioz Stars, 2026-04
90/100 stars
  Buy from Supplier
7 methoxy 1 methyl 9h pyrido 3 4 b indole  (Chem Impex International)
96
Chem Impex International 7 methoxy 1 methyl 9h pyrido 3 4 b indole
(A) ARS staining of mineralization in AS MSCs transduced with shHLA-B or shCtrl under osteogenic induction with quantification (B). (C) Immunoblot analyses showing the expression of RARB and TNAP in AS MSCs expressing shHLA-B or shCtrl at day 7 under osteogenic induction. (D) Immunoblot showing <t>HLA-B27</t> expressions of control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. (E) ARS staining of control MSCs transduced with control lentiviral vector (pLAS2w) or a vector expressing HLA-B27 (pLAS2w-HLA-B27) with quantification (F). (G) Immunoblot analyses showing RARB and TNAP expressions in control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. All experiments done in the AS patient group and the control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or 1-way ANOVA, followed by Tukey’s HSD test. Representative images from AS (A1) MSCs are shown in C. Scale bars: 200 μm (A and E).
7 Methoxy 1 Methyl 9h Pyrido 3 4 B Indole, supplied by Chem Impex International, 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/7 methoxy 1 methyl 9h pyrido 3 4 b indole/product/Chem Impex International
Average 96 stars, based on 1 article reviews
7 methoxy 1 methyl 9h pyrido 3 4 b indole - by Bioz Stars, 2026-04
96/100 stars
  Buy from Supplier
chem impex international u s a single  (Chem Impex International)
95
Chem Impex International chem impex international u s a single
(A) ARS staining of mineralization in AS MSCs transduced with shHLA-B or shCtrl under osteogenic induction with quantification (B). (C) Immunoblot analyses showing the expression of RARB and TNAP in AS MSCs expressing shHLA-B or shCtrl at day 7 under osteogenic induction. (D) Immunoblot showing <t>HLA-B27</t> expressions of control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. (E) ARS staining of control MSCs transduced with control lentiviral vector (pLAS2w) or a vector expressing HLA-B27 (pLAS2w-HLA-B27) with quantification (F). (G) Immunoblot analyses showing RARB and TNAP expressions in control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. All experiments done in the AS patient group and the control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or 1-way ANOVA, followed by Tukey’s HSD test. Representative images from AS (A1) MSCs are shown in C. Scale bars: 200 μm (A and E).
Chem Impex International U S A Single, supplied by Chem Impex International, 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/chem impex international u s a single/product/Chem Impex International
Average 95 stars, based on 1 article reviews
chem impex international u s a single - by Bioz Stars, 2026-04
95/100 stars
  Buy from Supplier
d l benzoyl phenylalanine  (Chem Impex International)
95
Chem Impex International d l benzoyl phenylalanine
(A) ARS staining of mineralization in AS MSCs transduced with shHLA-B or shCtrl under osteogenic induction with quantification (B). (C) Immunoblot analyses showing the expression of RARB and TNAP in AS MSCs expressing shHLA-B or shCtrl at day 7 under osteogenic induction. (D) Immunoblot showing <t>HLA-B27</t> expressions of control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. (E) ARS staining of control MSCs transduced with control lentiviral vector (pLAS2w) or a vector expressing HLA-B27 (pLAS2w-HLA-B27) with quantification (F). (G) Immunoblot analyses showing RARB and TNAP expressions in control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. All experiments done in the AS patient group and the control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or 1-way ANOVA, followed by Tukey’s HSD test. Representative images from AS (A1) MSCs are shown in C. Scale bars: 200 μm (A and E).
D L Benzoyl Phenylalanine, supplied by Chem Impex International, 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/d l benzoyl phenylalanine/product/Chem Impex International
Average 95 stars, based on 1 article reviews
d l benzoyl phenylalanine - by Bioz Stars, 2026-04
95/100 stars
  Buy from Supplier
fmoc  (Chem Impex International)
95
Chem Impex International fmoc
(A) ARS staining of mineralization in AS MSCs transduced with shHLA-B or shCtrl under osteogenic induction with quantification (B). (C) Immunoblot analyses showing the expression of RARB and TNAP in AS MSCs expressing shHLA-B or shCtrl at day 7 under osteogenic induction. (D) Immunoblot showing <t>HLA-B27</t> expressions of control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. (E) ARS staining of control MSCs transduced with control lentiviral vector (pLAS2w) or a vector expressing HLA-B27 (pLAS2w-HLA-B27) with quantification (F). (G) Immunoblot analyses showing RARB and TNAP expressions in control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. All experiments done in the AS patient group and the control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or 1-way ANOVA, followed by Tukey’s HSD test. Representative images from AS (A1) MSCs are shown in C. Scale bars: 200 μm (A and E).
Fmoc, supplied by Chem Impex International, 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/fmoc/product/Chem Impex International
Average 95 stars, based on 1 article reviews
fmoc - by Bioz Stars, 2026-04
95/100 stars
  Buy from Supplier
mouse monoclonal anti αb crystallin  (StressMarq)
90
StressMarq mouse monoclonal anti αb crystallin
(A) ARS staining of mineralization in AS MSCs transduced with shHLA-B or shCtrl under osteogenic induction with quantification (B). (C) Immunoblot analyses showing the expression of RARB and TNAP in AS MSCs expressing shHLA-B or shCtrl at day 7 under osteogenic induction. (D) Immunoblot showing <t>HLA-B27</t> expressions of control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. (E) ARS staining of control MSCs transduced with control lentiviral vector (pLAS2w) or a vector expressing HLA-B27 (pLAS2w-HLA-B27) with quantification (F). (G) Immunoblot analyses showing RARB and TNAP expressions in control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. All experiments done in the AS patient group and the control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or 1-way ANOVA, followed by Tukey’s HSD test. Representative images from AS (A1) MSCs are shown in C. Scale bars: 200 μm (A and E).
Mouse Monoclonal Anti αb Crystallin, supplied by StressMarq, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse monoclonal anti αb crystallin/product/StressMarq
Average 90 stars, based on 1 article reviews
mouse monoclonal anti αb crystallin - by Bioz Stars, 2026-04
90/100 stars
  Buy from Supplier
clotrimazole  (Chem Impex International)
95
Chem Impex International clotrimazole
(A) ARS staining of mineralization in AS MSCs transduced with shHLA-B or shCtrl under osteogenic induction with quantification (B). (C) Immunoblot analyses showing the expression of RARB and TNAP in AS MSCs expressing shHLA-B or shCtrl at day 7 under osteogenic induction. (D) Immunoblot showing <t>HLA-B27</t> expressions of control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. (E) ARS staining of control MSCs transduced with control lentiviral vector (pLAS2w) or a vector expressing HLA-B27 (pLAS2w-HLA-B27) with quantification (F). (G) Immunoblot analyses showing RARB and TNAP expressions in control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. All experiments done in the AS patient group and the control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or 1-way ANOVA, followed by Tukey’s HSD test. Representative images from AS (A1) MSCs are shown in C. Scale bars: 200 μm (A and E).
Clotrimazole, supplied by Chem Impex International, 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/clotrimazole/product/Chem Impex International
Average 95 stars, based on 1 article reviews
clotrimazole - by Bioz Stars, 2026-04
95/100 stars
  Buy from Supplier

Image Search Results


Canonical members of HSPB family prevent the aggregation of Aβ-GFP. Cells were transfected with HSPB1, HSPB5, HSPB7 or FRTTO at 1:3 ratio for 48 h. Quantification of the pellet/soluble ratio of Aβ-GFP relative to FRTTO was depicted in the chart above the blot. Values represent mean ± SE of two independent experiments. ** P < 0.01, ns = non significant.

Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of the amyloid beta-GFP fusion protein as a model of amyloid beta peptides-mediated aggregation: a study of DNAJB6 chaperone

doi: 10.3389/fnmol.2015.00040

Figure Lengend Snippet: Canonical members of HSPB family prevent the aggregation of Aβ-GFP. Cells were transfected with HSPB1, HSPB5, HSPB7 or FRTTO at 1:3 ratio for 48 h. Quantification of the pellet/soluble ratio of Aβ-GFP relative to FRTTO was depicted in the chart above the blot. Values represent mean ± SE of two independent experiments. ** P < 0.01, ns = non significant.

Article Snippet: The membranes were blocked with 5% dry milk in PBS with 0.1% Tween 20 (PBST) for 1 h at room temperature and incubated overnight at 4°C with the following primary antibodies: 6E10 (1:1000 in TBST, Covance), anti V5 (1:5000 in PBST, Invitrogen), anti β-actin (1:1000 in PBST, Abcam), anti HSPB1 (1:1000 in PBST, Stress Marq Biosciences), anti HSPB5 (1:1000 in PBST, Stress Marq Biosciences) and anti HSPB7 (1:1000, Abnova).

Techniques: Transfection

(A) Thiomaltose trimethoprim (TM-TMP): A new trimethoprim prodrug with enhanced water solubility that targets bacteria. (B) Proposed cleavage of TM-TMP in the presence of exogenous thiols.

Journal: Bioconjugate chemistry

Article Title: A Trimethoprim Conjugate of Thiomaltose Has Enhanced Antibacterial Efficacy In Vivo

doi: 10.1021/acs.bioconjchem.8b00177

Figure Lengend Snippet: (A) Thiomaltose trimethoprim (TM-TMP): A new trimethoprim prodrug with enhanced water solubility that targets bacteria. (B) Proposed cleavage of TM-TMP in the presence of exogenous thiols.

Article Snippet: Control groups included mice untreated with any drug, and mice treated with trimethoprim (Chem-Impex International Inc. Wood Dale, IL), while the experimental group was treated with thiomaltose-trimethoprim (TM-TMP).

Techniques: Solubility, Bacteria

Synthesis of thiomaltose-trimethoprim (TM-TMP). (a) DCM, TEA, rt, 55%. (b) DCC, DMAP, TEA, DMF, rt, 29%. (c) CuI, DIPEA, DMSO, rt, 65%.

Journal: Bioconjugate chemistry

Article Title: A Trimethoprim Conjugate of Thiomaltose Has Enhanced Antibacterial Efficacy In Vivo

doi: 10.1021/acs.bioconjchem.8b00177

Figure Lengend Snippet: Synthesis of thiomaltose-trimethoprim (TM-TMP). (a) DCM, TEA, rt, 55%. (b) DCC, DMAP, TEA, DMF, rt, 29%. (c) CuI, DIPEA, DMSO, rt, 65%.

Article Snippet: Control groups included mice untreated with any drug, and mice treated with trimethoprim (Chem-Impex International Inc. Wood Dale, IL), while the experimental group was treated with thiomaltose-trimethoprim (TM-TMP).

Techniques:

(A) ARS staining of mineralization in AS MSCs transduced with shHLA-B or shCtrl under osteogenic induction with quantification (B). (C) Immunoblot analyses showing the expression of RARB and TNAP in AS MSCs expressing shHLA-B or shCtrl at day 7 under osteogenic induction. (D) Immunoblot showing HLA-B27 expressions of control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. (E) ARS staining of control MSCs transduced with control lentiviral vector (pLAS2w) or a vector expressing HLA-B27 (pLAS2w-HLA-B27) with quantification (F). (G) Immunoblot analyses showing RARB and TNAP expressions in control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. All experiments done in the AS patient group and the control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or 1-way ANOVA, followed by Tukey’s HSD test. Representative images from AS (A1) MSCs are shown in C. Scale bars: 200 μm (A and E).

Journal: The Journal of Clinical Investigation

Article Title: HLA-B27–mediated activation of TNAP phosphatase promotes pathogenic syndesmophyte formation in ankylosing spondylitis

doi: 10.1172/JCI125212

Figure Lengend Snippet: (A) ARS staining of mineralization in AS MSCs transduced with shHLA-B or shCtrl under osteogenic induction with quantification (B). (C) Immunoblot analyses showing the expression of RARB and TNAP in AS MSCs expressing shHLA-B or shCtrl at day 7 under osteogenic induction. (D) Immunoblot showing HLA-B27 expressions of control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. (E) ARS staining of control MSCs transduced with control lentiviral vector (pLAS2w) or a vector expressing HLA-B27 (pLAS2w-HLA-B27) with quantification (F). (G) Immunoblot analyses showing RARB and TNAP expressions in control MSCs transduced with pLAS2w or pLAS2w-HLA-B27. All experiments done in the AS patient group and the control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or 1-way ANOVA, followed by Tukey’s HSD test. Representative images from AS (A1) MSCs are shown in C. Scale bars: 200 μm (A and E).

Article Snippet: The primary antibodies were mouse anti–active dephosphorylated β-catenin (clone 8E7; 1:500; Merck Millipore), rabbit anti–phosphorylated (serine 463/465) Smad 1/5/8 (clone 41D10; 1:250; Cell Signaling Technology), rabbit anti-TNAP (clone EPR4477; 1:500; Abcam), rabbit anti-RARB (catalog ab53161; 1:500; Abcam), goat anti-LEF1 (clone N-17; 1:500; Santa Cruz Biotechnology), rabbit anti-IRE1 (catalog ab37073; 1:1000; Abcam), rabbit anti–phosphorylated (serine 724) IRE1 (catalog ab4818; 1:500; Abcam), rabbit anti-XBP1 (catalog ab37152; 1:2000; Abcam), anti–HLA-B27 (clone HLA-ABC-m3; 1:300; Origene), anti–HLA-B7 [clone EPR2623( 2 ); 1:500; Abcam], rabbit anti-GAPDH (catalog GTX100118; 1:10,000; GeneTex), and mouse anti–β-actin (catalog MA5-15739; 1:10,000; Thermo Fisher Scientific).

Techniques: Staining, Transduction, Western Blot, Expressing, Plasmid Preparation

(A) Unfolded or misfolded HCs were immunoprecipitated (IP) from lysates of AS MSCs and control MSCs with HC10 antibody, followed by immunoblotting with HLA-B27 and anti-BiP antibodies. In AS MSCs, monomers of HLA-B27 HCs and dimers of disulfide-linked HLA-B27 HCs are indicated by an arrow and an arrowhead, respectively. (B and C) Immunoblot showing the expressions of p-IRE1 (B) and sXBP1 (C) protein in AS and control MSCs at day 7 under osteogenic induction. (D) Immunoblot showing the expressions of p-IRE1 and sXBP1 in AS MSCs transduced with shHLA-B or shCtrl. (E) Immunoblot showing the expressions of p-IRE1 and sXBP1 protein in control MSCs transduced with pLAS2w-HLA-B27 or pLAS2w. (F) ChIP assay showing sXBP1 binding at fragments (Frags) 1, 2, 3, 6, and 7 within 1400 bp upstream of the RARB transcriptional start site in AS MSCs. RARB 3′-untranslated region (3′UTR) was used as the negative control locus. (G) Immunoblot showing the expressions of RARB and TNAP in AS MSCs expressing shXBP1 or shCtrl. (H) ARS staining of mineralization in AS MSCs transduced with shXBP1 or shCtrl under osteogenic induction with quantification (I). All experiments done in the AS patient group and control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. **P < 0.05; ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or ****P < 0.0001 by 1-way ANOVA, followed by Tukey’s HSD test. Representative immunoblots from AS (A1) MSCs are shown in G.

Journal: The Journal of Clinical Investigation

Article Title: HLA-B27–mediated activation of TNAP phosphatase promotes pathogenic syndesmophyte formation in ankylosing spondylitis

doi: 10.1172/JCI125212

Figure Lengend Snippet: (A) Unfolded or misfolded HCs were immunoprecipitated (IP) from lysates of AS MSCs and control MSCs with HC10 antibody, followed by immunoblotting with HLA-B27 and anti-BiP antibodies. In AS MSCs, monomers of HLA-B27 HCs and dimers of disulfide-linked HLA-B27 HCs are indicated by an arrow and an arrowhead, respectively. (B and C) Immunoblot showing the expressions of p-IRE1 (B) and sXBP1 (C) protein in AS and control MSCs at day 7 under osteogenic induction. (D) Immunoblot showing the expressions of p-IRE1 and sXBP1 in AS MSCs transduced with shHLA-B or shCtrl. (E) Immunoblot showing the expressions of p-IRE1 and sXBP1 protein in control MSCs transduced with pLAS2w-HLA-B27 or pLAS2w. (F) ChIP assay showing sXBP1 binding at fragments (Frags) 1, 2, 3, 6, and 7 within 1400 bp upstream of the RARB transcriptional start site in AS MSCs. RARB 3′-untranslated region (3′UTR) was used as the negative control locus. (G) Immunoblot showing the expressions of RARB and TNAP in AS MSCs expressing shXBP1 or shCtrl. (H) ARS staining of mineralization in AS MSCs transduced with shXBP1 or shCtrl under osteogenic induction with quantification (I). All experiments done in the AS patient group and control group are from AS MSCs (A1, A2, and A3) and control MSCs (C1, C2, and C3), respectively, with at least 2–3 experimental repeats. Data are the mean ± SEM. **P < 0.05; ***P < 0.001; ****P < 0.0001 by 2-tailed Student’s t test (2 groups) or ****P < 0.0001 by 1-way ANOVA, followed by Tukey’s HSD test. Representative immunoblots from AS (A1) MSCs are shown in G.

Article Snippet: The primary antibodies were mouse anti–active dephosphorylated β-catenin (clone 8E7; 1:500; Merck Millipore), rabbit anti–phosphorylated (serine 463/465) Smad 1/5/8 (clone 41D10; 1:250; Cell Signaling Technology), rabbit anti-TNAP (clone EPR4477; 1:500; Abcam), rabbit anti-RARB (catalog ab53161; 1:500; Abcam), goat anti-LEF1 (clone N-17; 1:500; Santa Cruz Biotechnology), rabbit anti-IRE1 (catalog ab37073; 1:1000; Abcam), rabbit anti–phosphorylated (serine 724) IRE1 (catalog ab4818; 1:500; Abcam), rabbit anti-XBP1 (catalog ab37152; 1:2000; Abcam), anti–HLA-B27 (clone HLA-ABC-m3; 1:300; Origene), anti–HLA-B7 [clone EPR2623( 2 ); 1:500; Abcam], rabbit anti-GAPDH (catalog GTX100118; 1:10,000; GeneTex), and mouse anti–β-actin (catalog MA5-15739; 1:10,000; Thermo Fisher Scientific).

Techniques: Immunoprecipitation, Western Blot, Transduction, Binding Assay, Negative Control, Expressing, Staining