goat anti type iii collagen  (SouthernBiotech)

Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: FAM134B Controls Collagen I Dynamics in Hepatic Stellate Cell-Driven Fibrosis

doi: 10.1152/ajpgi.00170.2025

Figure Lengend Snippet: (a) Cas9 scramble, sgCCPG1, and sgFAM134B I and II cells were collected for immunoblot. Left panel shows quantitative immunoblots, and right panel shows quantifications. N = 3 biological replicates. (b, e) Cas9 scramble, sgCCPG1, and sgFAM134B I and II cells were treated with TGFβ (5 ng/mL) for 24 hours, and mRNA was collected for qRT-PCR. N = 6 biological replicates for (b), and N = 3 biological replicates for (e). (c) Cas9 scramble, sgCCPG1, and sgFAM134B I and II cells were treated with TGFβ (5 ng/mL) for 24 hours and collected for immunoblot to measure intracellular collagen I protein levels. Top panel shows representative immunoblots, and bottom panel shows quantifications. N = 3 biological replicates. (d) Cas9 scramble, sgCCPG1, and sgFAM134B II cells were treated with TGFβ (5 ng/mL) for 24 hours, and intracellular collagen I and LAMPI were stained with immunofluorescence of permeabilized cells. Top panel shows representative images (scale bars = 10μm for left panels and 2.5μm for the insets), and bottom panel shows quantifications. Samples were imaged by confocal microscopy at 63x with at least 10 images per sample, and colocalization was compared with a two-way ANOVA followed by a Tukey post-hoc. N = 5 biological replicates. (f) siControl, siCCPG1, and siFAM134B cells were treated with TGFβ (5 ng/mL) for 24 hours, and mRNA was collected for qRT-PCR. N = 4 biological replicates. (g) siControl, siCCPG1, and siFAM134B cells were treated with TGFβ (5 ng/mL) for 24 hours and collected for immunoblotting. Left panel shows representative immunoblots, and right panel shows quantifications. N = 3 biological replicates. Samples were analyzed with a one-way ANOVA (a) or two-way ANOVA (b-g) followed by a Tukey post-hoc. Graph bars represent sample mean and error bars depict standard deviation; ns = not significant, * = p < 0.05, ** = p < 0.01, and *** = p < 0.001.

Article Snippet: Antibodies used were as follows: Collagen I (Southern Biotech 1310–01), Collagen 5A1 (Cell Signaling Technologies 37304, RRID:AB_3675529), LAMP1 (Cell Signaling Technologies D2D11, RRID:AB_2687579), and Procollagen I (Millipore MAB1912, RRID:AB_94405).

Techniques: Western Blot, Quantitative RT-PCR, Staining, Immunofluorescence, Confocal Microscopy, Standard Deviation

Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: FAM134B Controls Collagen I Dynamics in Hepatic Stellate Cell-Driven Fibrosis

doi: 10.1152/ajpgi.00170.2025

Figure Lengend Snippet: (a) Cas9 scramble and sgFAM134B I and II cells were treated with TGFβ (5 ng/mL) for 24 hours, and collagen I deposition was stained with immunofluorescence of non-permeabilized cells. Left panel shows representative images (scale bar = 50μm), and right panel shows quantifications. Samples were imaged by confocal microscopy at 10x with at least 10 images per sample. Cas9 Control, sgCCPG1, and sgFAM134B I and II samples were compared with a two-way ANOVA followed by a Tukey post-hoc. N = 9 biological replicates. (b) LX-2 cells were transfected with scramble siRNA, siRNA against CCPG1, or siRNA against FAM134B (siControl, siCCPG1, and siFAM134B cells), and cells were treated with TGFβ (5 ng/mL) for 24 hours. Collagen I deposition was stained with immunofluorescence of non-permeabilized cells. Left panel shows representative images (scale bars = 30μm), and right panel shows quantifications. Samples were imaged by confocal microscopy at 20x with at least 10 images per sample. N = 7 biological replicates. (c) Cas9 scramble, sgCCPG1, and sgFAM134B I and II cells were treated with TGFβ (5 ng/mL) for 24 hours, and conditioned media samples were collected and concentrated for immunoblot. Left panel shows representative immunoblots, and right panel shows quantifications. N = 5 biological replicates. (d). LX-2 cells were treated with TGFβ (2ng/mL) for 16 hours, followed by CHX (50μM) ± CQ treatment (50μM). Conditioned media was harvested 4, 6, or 8 hours following CHX ± CQ addition and analyzed by immunoblotting. Coomassie staining for total protein serves as a control. Samples were analyzed with a two-way ANOVA followed by a Tukey post-hoc. Graph bars represent sample mean and error bars depict standard deviation; ns = not significant, * = p < 0.05, ** = p < 0.01, and *** = p < 0.001.

Article Snippet: Antibodies used were as follows: Collagen I (Southern Biotech 1310–01), Collagen 5A1 (Cell Signaling Technologies 37304, RRID:AB_3675529), LAMP1 (Cell Signaling Technologies D2D11, RRID:AB_2687579), and Procollagen I (Millipore MAB1912, RRID:AB_94405).

Techniques: Staining, Immunofluorescence, Confocal Microscopy, Control, Transfection, Western Blot, Standard Deviation

Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: FAM134B Controls Collagen I Dynamics in Hepatic Stellate Cell-Driven Fibrosis

doi: 10.1152/ajpgi.00170.2025

Figure Lengend Snippet: (a, b) Cas9 scramble, sgCCPG1, and sgFAM134B I and II cells were treated with TGFβ (5 ng/mL) for 24 hours, and intracellular collagen I (a) or procollagen I (b) were stained with immunofluorescence of permeabilized cells. Left panel shows representative images (scale bars = 10μm for left panels and 2.5μm for the insets), and right panel shows quantifications. Samples were imaged by confocal microscopy at 63x with at least 10 images per sample. Left panel shows representative images, and right panel shows quantifications. N = 7 biological replicates for (a), and N = 6 biological replicates for (b). Samples were analyzed with a two-way ANOVA followed by a Tukey post-hoc. Graph bars represent sample mean and error bars depict standard deviation; ns = not significant, * = p < 0.05 and ** = p < 0.01.

Article Snippet: Antibodies used were as follows: Collagen I (Southern Biotech 1310–01), Collagen 5A1 (Cell Signaling Technologies 37304, RRID:AB_3675529), LAMP1 (Cell Signaling Technologies D2D11, RRID:AB_2687579), and Procollagen I (Millipore MAB1912, RRID:AB_94405).

Techniques: Staining, Immunofluorescence, Confocal Microscopy, Standard Deviation

Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: FAM134B Controls Collagen I Dynamics in Hepatic Stellate Cell-Driven Fibrosis

doi: 10.1152/ajpgi.00170.2025

Figure Lengend Snippet: (a) During HSC activation, COL1A1 mRNA is transcribed in the nucleus and translated into the ER for folding. Misfolded collagen I aggregates are trafficked to the lysosome for degradation through ER-phagy mediated by FAM134B, while folded collagen I is secreted from the cell into the extracellular space for deposition into collagen fibrils. (b) The loss of FAM134B prevents trafficking of misfolded collagen I aggregates to the lysosome. To restore ER homeostasis, we predict that misfolded collagen I is secreted from the cell, where it interferes with normal collagen fibril assembly and deposition. Created in BioRender. Maiers, J. (2025) https://BioRender.com/lu8v4nq .

Article Snippet: Antibodies used were as follows: Collagen I (Southern Biotech 1310–01), Collagen 5A1 (Cell Signaling Technologies 37304, RRID:AB_3675529), LAMP1 (Cell Signaling Technologies D2D11, RRID:AB_2687579), and Procollagen I (Millipore MAB1912, RRID:AB_94405).

Techniques: Activation Assay