Journal: Bioactive Materials

Article Title: Bioinspired scaffold recapitulating chondrogenic ontogeny and microenvironment for functional cartilage regeneration

doi: 10.1016/j.bioactmat.2025.11.041

Figure Lengend Snippet: Biological functional analysis of tri-layer scaffold systems. A) Representative images of migrated cells stained with crystal violet at 48 h. B) Number of migrating cells in control and CE-SKP. C) Glycosaminoglycan concentrations in control and CE-SKP at 7 and 14 days. D) qPCR analysis for COL2, SOX9, and ACAN. E) Western blotting of COL2 and SOX9. F) Alizarin red staining of control and CPH. G) ALP activity of control and CPH. H) qPCR analysis for COL1, COL10, OCN, and RUNX2. I) Western blotting of COL10, COL1, and RUNX2. J) Representative images of migrated cells stained with crystal violet at 48 h. K) Number of migrating cells in control and P2G3. L) Scanning electron microscope micrographs of P2G3. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: To evaluate the regeneration of cartilage, COL1 (1:100, HY- P81227 , MCE), COL2 (1:100, ARG20787 , Arigobio), and COL10 (1:100, BA2023, Boster) were used as a marker in immunohistochemistry (IHC) staining.

Techniques: Functional Assay, Staining, Control, Western Blot, Activity Assay, Microscopy

Journal: Bioactive Materials

Article Title: Bioinspired scaffold recapitulating chondrogenic ontogeny and microenvironment for functional cartilage regeneration

doi: 10.1016/j.bioactmat.2025.11.041

Figure Lengend Snippet: Regeneration of the femoral trochlea in a rabbit model. Representative images of IHC staining for A) COL1, B) COL10.

Article Snippet: To evaluate the regeneration of cartilage, COL1 (1:100, HY- P81227 , MCE), COL2 (1:100, ARG20787 , Arigobio), and COL10 (1:100, BA2023, Boster) were used as a marker in immunohistochemistry (IHC) staining.

Techniques: Immunohistochemistry

Journal: Journal of Clinical Investigation

Article Title: Endoplasmic reticulum protein TXNDC5 promotes renal fibrosis by enforcing TGF-β signaling in kidney fibroblasts

doi: 10.1172/jci143645

Figure Lengend Snippet: Figure 2. TXNDC5 was highly upregulated in renal fibroblasts, but not in TECs, endothelial cells, or podocytes, of the fibrotic kidneys. (A and C) IF stain- ing of TXNDC5 (red) on sections of fibrotic kidneys induced by UUO in Col1a1-GFPTg mice showed TXNDC5 was mainly expressed in collagen-secreting renal fibroblasts (green), both in renal cortex and medulla (n = 6). Cell nuclei were stained with DAPI (blue). Scale bar: 100 μm. (B) IF staining of TXNDC5 (green) on section of fibrotic kidneys induced by UUO in Cdh16-Cre, NPHS2-Cre, and Tie2-Cre/ERT2 tdTomato mice. Cell nuclei were stained with DAPI (blue). Scale bar: 100 μm. (D) A pie chart to illustrate the proportion of TXNDC5+ cells in different types of kidney cells in the UUO-induced fibrotic kidneys. Data are representative of 3 or more independent experimental replicates. Data in C are presented as mean ± SEM.

Article Snippet: COL1A1 (1:1000, Aviva Systems Biology, OAAB10798, for mouse species), COL1A1 (1:1000, OriGene, TA309096, for human species), CCN2, TGFBR2 (1:1000, OriGene, TA323092, TA311643), POSTN, ATF6, N-cadherin (1:1000, 1:1000, 1:3000, GeneTex, GTX100602, GTX104820, GTX127345), FN1 (1:1000, BD Biosciences, 610077), TXNDC5, β-tubulin (1:30000, 1:5000, Proteintech, 19834-1-AP, 66240-1-Ig), BiP (1:1000, Cell Signaling Technology, 3177), p-SMAD3 (1:1000, Abcam, ab40854), totalSMAD3 (1:1000, Cell Signaling Technology, 9523, for mouse species), total-SMAD3 (1:1000, Abcam, ab52903, for human species), TGFBR1 (1:1000, Thermo Fisher Scientific, PA5-32631, for human species), TGFBR1 (1:1000, Abcam, ab31013, for mouse species), and GAPDH (1:5000, Thermo Fisher Scientific, MA5-15738).

Techniques: Staining

Journal: Journal of Clinical Investigation

Article Title: Endoplasmic reticulum protein TXNDC5 promotes renal fibrosis by enforcing TGF-β signaling in kidney fibroblasts

doi: 10.1172/jci143645

Figure Lengend Snippet: Figure 3. Knockdown of TXNDC5 attenuated TGF-β1–induced HKF activation and ECM production; overexpression of TXNDC5 was sufficient to trigger HKF activation and ECM production. (A) Protein and (B) transcript expression levels of fibroblast activation marker (periostin) and ECM proteins (COL1A1, fibronectin, and CCN2) were increased in control (Scramble) HKFs following TGF-β1 (10 ng/mL) treatment. Knockdown of TXNDC5 attenuated the upregu- lation of these fibrogenic markers induced by TGF-β1 in HKFs (n = 5–10). (C) Overexpression of TXNDC5 was sufficient to induce upregulation of fibroblast activation marker (Periostin) and ECM proteins (COL1A1, fibronectin) in HKFs (n = 3–10). (D) Treatment of TGF-β1 (10 ng/mL) increased the cellular prolifera- tion activity of HKFs, which was abrogated by TXNDC5 knockdown. (E) Overexpression of TXNDC5 increased the cellular proliferation activity of HKFs. In D and E, n = 10. Data are representative of 3 or more independent experimental replicates. For all panels, data are presented as mean ± SEM. The statistical significance of differences for 2 groups was determined by 2-sided t test and among 3 or more groups it was determined using 1-way ANOVA, followed by Sidak’s post hoc tests. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: COL1A1 (1:1000, Aviva Systems Biology, OAAB10798, for mouse species), COL1A1 (1:1000, OriGene, TA309096, for human species), CCN2, TGFBR2 (1:1000, OriGene, TA323092, TA311643), POSTN, ATF6, N-cadherin (1:1000, 1:1000, 1:3000, GeneTex, GTX100602, GTX104820, GTX127345), FN1 (1:1000, BD Biosciences, 610077), TXNDC5, β-tubulin (1:30000, 1:5000, Proteintech, 19834-1-AP, 66240-1-Ig), BiP (1:1000, Cell Signaling Technology, 3177), p-SMAD3 (1:1000, Abcam, ab40854), totalSMAD3 (1:1000, Cell Signaling Technology, 9523, for mouse species), total-SMAD3 (1:1000, Abcam, ab52903, for human species), TGFBR1 (1:1000, Thermo Fisher Scientific, PA5-32631, for human species), TGFBR1 (1:1000, Abcam, ab31013, for mouse species), and GAPDH (1:5000, Thermo Fisher Scientific, MA5-15738).

Techniques: Knockdown, Activation Assay, Over Expression, Expressing, Marker, Control, Activity Assay

Journal: Journal of Clinical Investigation

Article Title: Endoplasmic reticulum protein TXNDC5 promotes renal fibrosis by enforcing TGF-β signaling in kidney fibroblasts

doi: 10.1172/jci143645

Figure Lengend Snippet: Figure 9. Targeted deletion of Txndc5 in renal fibroblasts attenuated kidney fibrosis. (A) Illustration of experimental design to induce Txndc5 deletion specifically in renal fibroblasts. (B) Picrosirius red staining of kidney sections from WT and Txndc5cKO mice 10 days after UUO (n = 6–7). Scale bar: 50 μm. (C) Immunoblots to quantify fibroblast activation marker (POSTN) and ECM (COL1A1 and CCN2) proteins in whole-kidney lysates from Col1a2-Cre and Txndc5cKO mice 10 days after UUO (n = 5–6). (D) SHG images of kidney sections from Col1a2-Cre and Txndc5cKO mice 10 days after UUO. The quantitative results of SHG-positive areas showed accumulation of fibrillar collagen in Col1a2-Cre kidneys, which was ameliorated in Txndc5cKO mice (n = 3). Scale bar: 50 μm. Data are representative of 3 or more independent experimental replicates. For all panels, data are presented as mean ± SEM. The statistical significance of differences among 3 or more groups was determined using 1-way ANOVA, followed by Sidak’s post hoc tests. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: COL1A1 (1:1000, Aviva Systems Biology, OAAB10798, for mouse species), COL1A1 (1:1000, OriGene, TA309096, for human species), CCN2, TGFBR2 (1:1000, OriGene, TA323092, TA311643), POSTN, ATF6, N-cadherin (1:1000, 1:1000, 1:3000, GeneTex, GTX100602, GTX104820, GTX127345), FN1 (1:1000, BD Biosciences, 610077), TXNDC5, β-tubulin (1:30000, 1:5000, Proteintech, 19834-1-AP, 66240-1-Ig), BiP (1:1000, Cell Signaling Technology, 3177), p-SMAD3 (1:1000, Abcam, ab40854), totalSMAD3 (1:1000, Cell Signaling Technology, 9523, for mouse species), total-SMAD3 (1:1000, Abcam, ab52903, for human species), TGFBR1 (1:1000, Thermo Fisher Scientific, PA5-32631, for human species), TGFBR1 (1:1000, Abcam, ab31013, for mouse species), and GAPDH (1:5000, Thermo Fisher Scientific, MA5-15738).

Techniques: Staining, Western Blot, Activation Assay, Marker

Journal: Journal of Clinical Investigation

Article Title: Endoplasmic reticulum protein TXNDC5 promotes renal fibrosis by enforcing TGF-β signaling in kidney fibroblasts

doi: 10.1172/jci143645

Figure Lengend Snippet: Figure 10. Targeted deletion of Txndc5 in renal fibroblasts mitigated kidney fibrosis induced by uIRI. (A) Picrosirius red staining of kidney sections from WT and Txndc5cKO mice 28 days after uIRI (n = 7–11). Scale bar: 50 μm. (B) Immunoblots to quantify fibroblast activation marker (POSTN) and ECM (COL1A1 and CCN2) proteins in whole-kidney lysates from Col1a2-Cre and Txndc5cKO mice 28 days after uIRI (n = 4–11). (C) SHG images of kidney sections from Col1a2-Cre and Txndc5cKO mice 28 days after uIRI. The quantitative results of SHG-positive areas showed accumulation of fibrillar collagen in Col1a2-Cre kidneys, which was ameliorated in Txndc5cKO mice (n = 3). Scale bar: 50 μm. Data are representative of 3 or more independent experimental replicates. For all panels, data are presented as mean ± SEM. The statistical significance of differences among 3 or more groups was determined using 1-way ANOVA, followed by Sidak’s post hoc tests. **P < 0.01, ***P < 0.001.

Article Snippet: COL1A1 (1:1000, Aviva Systems Biology, OAAB10798, for mouse species), COL1A1 (1:1000, OriGene, TA309096, for human species), CCN2, TGFBR2 (1:1000, OriGene, TA323092, TA311643), POSTN, ATF6, N-cadherin (1:1000, 1:1000, 1:3000, GeneTex, GTX100602, GTX104820, GTX127345), FN1 (1:1000, BD Biosciences, 610077), TXNDC5, β-tubulin (1:30000, 1:5000, Proteintech, 19834-1-AP, 66240-1-Ig), BiP (1:1000, Cell Signaling Technology, 3177), p-SMAD3 (1:1000, Abcam, ab40854), totalSMAD3 (1:1000, Cell Signaling Technology, 9523, for mouse species), total-SMAD3 (1:1000, Abcam, ab52903, for human species), TGFBR1 (1:1000, Thermo Fisher Scientific, PA5-32631, for human species), TGFBR1 (1:1000, Abcam, ab31013, for mouse species), and GAPDH (1:5000, Thermo Fisher Scientific, MA5-15738).

Techniques: Staining, Western Blot, Activation Assay, Marker

Journal: Journal of Clinical Investigation

Article Title: Endoplasmic reticulum protein TXNDC5 promotes renal fibrosis by enforcing TGF-β signaling in kidney fibroblasts

doi: 10.1172/jci143645

Figure Lengend Snippet: Figure 11. Induced deletion of Txndc5 in renal fibroblasts mitigated the progression of established kidney fibrosis. (A) Illustration of experimental design to induce Txndc5 deletion specifically in renal fibroblasts in mouse kidneys with established fibrosis. (B) Picrosirius red staining of kidney sections from Col1a2-Cre and Txndc5cKO mice. Ten days after UUO, Col1a2-Cre and Txndc5cKO mice showed a similar extent of renal fibrosis prior to tamoxifen injec- tion. Eleven days after tamoxifen treatment, the fibrotic areas more than doubled (increased from 5.1% to 10.6%) in Col1a2-Cre, but barely changed in Txndc5cKO (changed from 5.3% to 6.9%) mouse kidneys (n = 5–6). Scale bar: 50 μm. (C) Protein expression levels of fibroblast activation marker (perios- tin), ECM (CCN2), and TGFBR1 in whole-kidney lysate from Col1a2-Cre and Txndc5cKO mice (n = 4–6). (D) Schematic summary of the proposed profibrotic mechanisms by which TXNDC5 contributes to the pathogenesis of renal fibrosis. TF: transcription factor. Data are representative of 3 or more independent experimental replicates. For all panels, data are presented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 by 2-sided t test.

Article Snippet: COL1A1 (1:1000, Aviva Systems Biology, OAAB10798, for mouse species), COL1A1 (1:1000, OriGene, TA309096, for human species), CCN2, TGFBR2 (1:1000, OriGene, TA323092, TA311643), POSTN, ATF6, N-cadherin (1:1000, 1:1000, 1:3000, GeneTex, GTX100602, GTX104820, GTX127345), FN1 (1:1000, BD Biosciences, 610077), TXNDC5, β-tubulin (1:30000, 1:5000, Proteintech, 19834-1-AP, 66240-1-Ig), BiP (1:1000, Cell Signaling Technology, 3177), p-SMAD3 (1:1000, Abcam, ab40854), totalSMAD3 (1:1000, Cell Signaling Technology, 9523, for mouse species), total-SMAD3 (1:1000, Abcam, ab52903, for human species), TGFBR1 (1:1000, Thermo Fisher Scientific, PA5-32631, for human species), TGFBR1 (1:1000, Abcam, ab31013, for mouse species), and GAPDH (1:5000, Thermo Fisher Scientific, MA5-15738).

Techniques: Staining, Expressing, Activation Assay, Marker

Journal: Biological Research

Article Title: Effect of cytoglobin overexpression on extracellular matrix component synthesis in human tenon fibroblasts

doi: 10.1186/s40659-019-0229-4

Figure Lengend Snippet: a RT-PCR analysis of mRNA expression levels of collagen I, collagen III and fibronectin in human tenon fibroblasts after Cygb overexpression. Data are represented as the mean ± standard deviation (n = 3), *p < 0.05; **p < 0.01, t test. b Western blot analysis of collagen I, collagen III, fibronectin and lamin B (as control) in human tenon fibroblasts after Cygb overexpression

Article Snippet: The membranes were blocked with 5% skim milkand incubated with primary antibodies including lamin B (1:1000; 66095-1-Ig; ProteinTech Group, Chicago, IL, USA), GAPDH (1:1000; M20005 M; Abmart, China), FLAG (1:1000; F1804; Sigma, St. Louis, MO, USA), hypoxia-inducible factor-1 alpha (HIF-1α) (1:1000; ab51608; Abcam, Cambridge, MA, USA), transforming growth factor (TGF)-β1 (1:1000; ab179695; Abcam), fibronectin (1:1000; ab32419; Abcam), collagen III (1:1000; BA0326; Boster, Wuhan, China), and collagen I (1:1000; BA0325; Boster).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Over Expression, Standard Deviation, Western Blot