Journal: iScience

Article Title: Human iPSC cardiomyocyte patch transplantation modifies extracellular matrix and fibroblast behavior after myocardial infarction

doi: 10.1016/j.isci.2026.115341

Figure Lengend Snippet: TGF-β secreted from the hiPSC-derived CM patch induced Col1a1 expression but not Col3a1 (A) Schematic representation of the co-culture system. hiPSC-derived CMs were seeded on the upper chamber, whereas the cardiac fibroblasts isolated from mice heart were cultured at the lower chamber in the presence and absence of SB431542, a TGF-β receptor inhibitor. (B) RT-qPCR analysis of Col1a1 , Col3a1 , and Pai1 in the cultured cardiac fibroblasts. Two-way ANOVA (co-culture × inhibitor) with interaction; Tukey-adjusted post hoc tests on estimated marginal means. Data represent mean ± SEM. N = 3–6, one-way ANOVA followed by Tukey’s HSD test, p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Then proteins were analyzed using primary antibodies for Col1a1 (R1038, OriGene, Rockville, MD), Col3a1(ab6310, Abcam, Cambridge, UK) and Gapdh (AM4300, Thermo Fisher Scienctific, Waltham, MA), respectively on Amersham Imager 600 (GE Healthcare, Chicago, IL).

Techniques: Derivative Assay, Expressing, Co-Culture Assay, Isolation, Cell Culture, Quantitative RT-PCR

Journal: The 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: (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 upregulation 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 proliferation 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), total-SMAD3 (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: Expressing, Activation Assay, Marker, Control, Knockdown, Over Expression, Activity Assay

Journal: The 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: (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), total-SMAD3 (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: The 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: (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), total-SMAD3 (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: The 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: (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 injection. 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 (periostin), 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), total-SMAD3 (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, Injection, Expressing, Activation Assay, Marker