anti collagen  (OriGene)

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 and C) IF staining 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), 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

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, 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) Picrosirius red staining (top 2 panels) and Masson’s trichrome staining (bottom 2 panels) of kidney sections from WT and Txndc5–/– mice 10 days after UUO. Bar graphs of the quantitative results of Picrosirius red and Masson’s trichrome staining areas are shown on the right (n = 5–10). Scale bar: 100 μm. (B) SHG images of the kidney sections from WT and Txndc5–/– mice 10 days after UUO. The quantitative results of SHG-positive areas showed increased accumulation of fibrillar collagen (green) in WT but not in Txndc5–/– mice kidneys following injury. For each of the kidney sections imaged for SHG, TPEF imaging was obtained to show the profile of the scanned tissue (red color in bottom panels) (n = 3). Scale bar: 50 μm. (C) Immunoblots showed protein expression levels of fibroblast activation marker (Periostin) and ECM (COL1A1) in the whole-kidney extract from WT and Txndc5–/– mice 10 days after UUO (n = 3–6). 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, Imaging, Western Blot, Expressing, 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 (top 2 panels) and Masson’s trichrome staining (bottom 2 panels) of kidney sections from WT and Txndc5–/– mice 28 days after uIRI. Bar graphs of the quantitative results of Picrosirius red and Masson’s trichrome staining areas are shown on the right (n = 6). Scale bar: 100 μm. (B) SHG images of the kidney sections from WT and Txndc5–/– mice 28 days after uIRI. The quantitative results of SHG-positive areas showed increased accumulation of fibrillar collagen (green) in WT but not in Txndc5–/– mice kidneys following injury. For each of the kidney sections imaged for SHG, TPEF imaging was obtained to show the profile of the scanned tissue (red color in bottom panels) (n = 3). Scale bar: 50 μm. (C) Immunoblots showed protein expression levels of fibroblast activation marker (Periostin) and ECM (COL1A1) in the whole-kidney extract from WT and Txndc5–/– mice 28 days after uIRI (n = 5–9). 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, Imaging, Western Blot, Expressing, 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. (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