Journal: Molecular metabolism

Article Title: Targeting hepatic oxidative stress rescues bone loss in liver fibrosis.

doi: 10.1016/j.molmet.2022.101599

Figure Lengend Snippet: Figure 4: Stanniocalcin 1 (STC1) knock-down attenuates the effects of SHED-Heps on CCl4-induced ROS production and cell survival of mouse hepatocytes (mHeps). SHED-Heps were treated with siRNA specific for STC1 (siSTC1) and scrambled control siRNA (siCONT), referred to as siSTC1-SHED-Heps and siCONT-SHED-Heps. (A) The graph presents the expression of STC1 in SHED-Heps by RT-qPCR. The results are presented as a ratio of the expression in siCONT-SHED-Heps (siCONT ¼ 1). (B) Representative images of STC1 in SHED-Heps were detected by immunofluorescence analysis. The nuclei were stained with DAPI. Scale bar, 30 mm. (C) The graph shows the levels of STC1 in the conditioned medium of SHED-Heps by ELISA. (D, E) mHeps were co-cultured with SHED-Heps under the stimulation of CCl4 (2 mg/mL). The graph presents the production of ROS (D) and cell viability (E) in mHeps by colorimetric analysis. The results are presented as a ratio of the expression in mHeps without CCl4 stimulation (w/o CCl4 ¼ 1). A, CeE: n ¼ 5 for all groups. *P ˂ 0.05, ***P ˂ 0.005. #P ˂ 0.05, ##P ˂ 0.01, ###P ˂ 0.005. (vs. CCl4-treated mHeps). ns, no significant difference (vs. CCl4-treated mHeps). The graph bars represent the mean SEM.

Article Snippet: SHED-Heps were pretreated for 3 days with small interfering RNA (siRNA) specific for stanniocalcin 1 (STC1) or with scrambled control (referred to as siSTC1 and siCONT, respectively; Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Knockdown, Control, Expressing, Quantitative RT-PCR, Staining, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Molecular metabolism

Article Title: Targeting hepatic oxidative stress rescues bone loss in liver fibrosis.

doi: 10.1016/j.molmet.2022.101599

Figure Lengend Snippet: Figure 5: STC1 knock-down attenuates the effects of SHED-HepTx on the anti-fibro-inflammation and ROS production in CCl4-treated mice. CCl4-treated mice were harvested four weeks after SHED-HepTx. (A) The graphs present the serum levels of AST, ALT, and total bilirubin by colorimetric analysis. (B, C) Representative liver images were analyzed by Sirius Red staining (B). Representative liver images of ACTA2 were acquired by immunohistochemical analysis. Nuclei were stained with hematoxylin (C). Scale bars: 200 mm (B) and 50 mm (C). PV: portal vein. (D) The graph presents the hepatic HYP content by colorimetric analysis. (E) The graphs present the hepatic expression of Col1a1 and Acta2 by RT-qPCR. (F) The graphs present the hepatic expression of Il-17 by RT-qPCR. (G) The graphs present the hepatic levels of MDA and GSH-Px livers by colorimetric analysis. (H) The graphs present the hepatic expression of Pparg and Nox4 by RT-qPCR. The results are presented as a ratio of the expression in the CCl4 group (CCl4 ¼ 1). AeH: CCl4, CCl4- treated group; SHED-HepTx, SHED-HepTx group; siCONT, siCONT treatment; siSTC1, siSTC1 treatment. A, DeH: n ¼ 10 in all groups. *P ˂ 0.05, **P ˂ 0.01, ***P ˂ 0.005. ns, no significance. The graph bars represent the mean SEM. E, F, H: The results are presented as a ratio of the expression in the CCl4-treated group (CCl4 ¼ 1).

Article Snippet: SHED-Heps were pretreated for 3 days with small interfering RNA (siRNA) specific for stanniocalcin 1 (STC1) or with scrambled control (referred to as siSTC1 and siCONT, respectively; Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Knockdown, Staining, Immunohistochemical staining, Expressing, Quantitative RT-PCR

Journal: Molecular metabolism

Article Title: Targeting hepatic oxidative stress rescues bone loss in liver fibrosis.

doi: 10.1016/j.molmet.2022.101599

Figure Lengend Snippet: Figure 6: STC1 knock-down attenuates the bone recovery in CCl4-treated mice with SHED-HepTx. CCl4-treated mice were harvested four weeks after SHED-HepTx. (AeD) Representative 2D (A) and 3D (B) images of proximal tibiae by microCT analysis. The graphs present BMD (C) and bone parameters (D). (E) The graphs present the serum levels of CTX-I and TRAP-5b by ELISA. AeE: CCl4, CCl4-treated group; SHED-HepTx, SHED-HepTx group; siCONT, siCONT treatment; siSTC1, siSTC1 treatment. CeE: n ¼ 10 for all groups. *P ˂ 0.05, **P ˂ 0.01, ***P ˂ 0.005. ns, no significance. The graph bars represent the mean SEM.

Article Snippet: SHED-Heps were pretreated for 3 days with small interfering RNA (siRNA) specific for stanniocalcin 1 (STC1) or with scrambled control (referred to as siSTC1 and siCONT, respectively; Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Knockdown, Enzyme-linked Immunosorbent Assay

Journal: Molecular metabolism

Article Title: Targeting hepatic oxidative stress rescues bone loss in liver fibrosis.

doi: 10.1016/j.molmet.2022.101599

Figure Lengend Snippet: Figure 7: STC1 knock-down attenuates the suppression of osteoclast differentiation via IL-17 enhanced tumor necrosis factor superfamily 11 (TNFSF11) in CCl4- treated mice with SHED-HepTx.BMCs were co-cultured with calvarial osteoblasts. (AeC) CCl4-treated mice were harvested four weeks after SHED-HepTx. Representative images of osteoclast differentiation were analyzed by TRAP staining (A). The graph presents the number of TRAPþ MNCs. (B). The graphs indicate the expression of Il-17 and Tnfrsf11a in BMCs according to RT-qPCR. The results are presented as a ratio to the expression in the CCl4 group (CCl4 ¼ 1) (C). (D, E) The co-cultures were treated with or without re- combinant mouse IL-17 (10 nM) and/or anti-mouse TNFSF11 goat IgG (TNFSF11 Ab; 50 ng/mL) or its control IgG antibody (Cont Ab). Representative images of osteoclast dif- ferentiation were analyzed by TRAP staining (D). The graph presents the number of TRAPþ MNCs. n ¼ 5 for all groups. ###P ˂ 0.005 vs. control group without IL-17, TNFSF11 Ab, and Cont Ab treatment NS, no significance vs. control group. (E). AeC: CCl4, CCl4-treated group; SHED-HepTx, SHED-HepTx group; siCONT, siCONT treatment; siSTC1, siSTC1 treatment. B, C, E: *P ˂ 0.05, **P ˂ 0.01, ***P ˂ 0.005. ns, no significance. The graph bars represent the mean SEM. B, C: n ¼ 10 for all groups.

Article Snippet: SHED-Heps were pretreated for 3 days with small interfering RNA (siRNA) specific for stanniocalcin 1 (STC1) or with scrambled control (referred to as siSTC1 and siCONT, respectively; Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Knockdown, Cell Culture, Staining, Expressing, Quantitative RT-PCR, Control

Journal: Bioelectricity

Article Title: Membrane Potential Depolarization Alters Calcium Flux and Phosphate Signaling During Osteogenic Differentiation of Human Mesenchymal Stem Cells

doi: 10.1089/bioe.2018.0005

Figure Lengend Snippet: STC1 gene expression is upregulated with depolarization, and STC1 knockdown rescues early OS differentiation markers, but not calcification, in depolarized hMSCs. STC1 expression (A) and sodium phosphate cotransporter SLC20A1 expression (B) were quantified on day 7 by qPCR in OS-differentiated cells cultured under depolarizing (HK) or nondepolarizing (OS) conditions with or without 0.5 mM Pi supplementation. (C) STC1 expression was quantified 24 h after STC1 knockdown by siRNA. STC1-knockdown cells and negative control knockdown cells were then cultured in OS medium, OS medium with high K+ (HK), or control medium (CON). On day 7, cultures were evaluated for expression of STC1 (D), ALPL (E), or IBSP (F). Data points are mean normalized expression ± standard deviation, n = 4–6. (G, H) On day 21, cultures were evaluated for total calcium content. Data points represent mean normalized calcium content (μg Ca/μg DNA) ± standard deviation, n = 4. Different letters over bar graphs represent statistically different groups as determined by one-way ANOVA and the Tukey–Kramer post hoc test, p < 0.05. Asterisks (*) represent statistical differences between STC1-knockdown versus negative control knockdown cells, as determined by the Student's t-test, p < 0.05. ALPL, alkaline phosphatase gene; STC1, Stanniocalcin 1.

Article Snippet: Lipofectamine RNAiMAX was used to transfect STC1-specific Silencer Select siRNA (Invitrogen). siRNA against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a positive control for transfection.

Techniques: Expressing, Cell Culture, Negative Control, Standard Deviation

Journal: Bioelectricity

Article Title: Membrane Potential Depolarization Alters Calcium Flux and Phosphate Signaling During Osteogenic Differentiation of Human Mesenchymal Stem Cells

doi: 10.1089/bioe.2018.0005

Figure Lengend Snippet: Schematic depicting proposed phosphate- and STC1-related interactions involved in hMSC depolarization during OS differentiation. Vmem depol may alter Na+ and Pi transport across the cell membrane through the Na+-dependent Pi cotransporter SLC20A1. Pi levels, in turn, may alter intracellular STC1 expression. STC1 regulates the expression of OS genes such as ALPL and IBSP and also critically regulates late-stage mineralization. Vmem depol, membrane potential depolarization.

Article Snippet: Lipofectamine RNAiMAX was used to transfect STC1-specific Silencer Select siRNA (Invitrogen). siRNA against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a positive control for transfection.

Techniques: Expressing