Journal: Mediators of Inflammation

Article Title: Endoplasmic Reticulum Stress in Colonic Mucosa of Ulcerative Colitis Patients Is Mediated by PERK and IRE1 Pathway Activation

doi: 10.1155/2022/6049500

Figure Lengend Snippet: Activation of UPR-responsive genes and chaperone modulation. (a) Transcriptional levels of STC2, ATF3, and DDIT3 are shown as a fold change; ∗ p < 0.05, ∗∗ p < 0.005. (b) Quantitative analysis of immunohistochemistry staining for DDIT3 of both groups. For UC, n = 6; for CTR, n = 3; ∗ p < 0.05. (c) Immunohistochemical analysis of DDIT3 was performed on paraffin-embedded slides from intestinal mucosa of both UC and CTR groups. The arrows indicate DDIT3-positive cells, which are shown in brown. The white arrows indicate positive epithelial cells, and the black arrows signalize positive cells from the lamina propria. Scale bars: 50 μ m. (d) Transcriptional levels of the chaperones DNAJC3, CALR, HSPA5, and HSP90B1 are shown as fold change, ∗∗ p < 0.005. (e, f) Immunohistochemical analysis of GRP78 and GRP94 was performed on paraffin-embedded slides from intestinal mucosa of both groups. The arrows indicate GRP78- and GRP94-positive cells, which are shown in brown. The white arrows indicate positive epithelial cells, and the black arrows signalize positive cells from the lamina propria. Scale bars: 50 μ m. (g, h) Quantitative analysis of immunohistochemistry staining for GRP78 and GRP94 of both groups. (g) For UC, n = 8; for CTR, n = 5; ∗∗ p < 0.005. (h) For UC, n = 6; for CTR, n = 4; ∗∗ p < 0.005. CTR = control; UC = ulcerative colitis.

Article Snippet: To evaluate UPR-related genes and chaperones, the primers used were ATF3 (Hs_00231069_m1), CALR (Hs_00189032_m1), STC2 (Hs_00175027_m1), DNAJC3 (Hs_00534483_m1), DDIT3 (Hs_0109850_m1), HSP90B1 (Hs_00427665_g1), and HSPA5 (Hs_99999174_m1).

Techniques: Activation Assay, Immunohistochemistry, Staining, Immunohistochemical staining, Control

Journal: Life Science Alliance

Article Title: Identification and characterization of a membrane receptor that binds to human STC1

doi: 10.26508/lsa.202201497

Figure Lengend Snippet: (A) hSTC1, (B) hSTC2, and (C) hCREG binding to human IGF2R.

Article Snippet: To perform kinetic affinity assays, hSTC1-His, human His-tagged STC2 (hSTC2-His, 9405-SO-050; R&D Systems), and human His-tagged cellular repressor of E1A-stimulated gene (hCREG-His, 2380-CR-025/CF; R&D Systems) were prepared at 10 μg/ml in a running buffer, followed by a twofold serial dilution to give 10, 5, 2.5, 1.25, 0.625, 0.313, 0.156, 0.078, 0.039, and 0.019 μg/ml solutions.

Techniques: Binding Assay

Journal: Neoplasma

Article Title: Blocking stanniocalcin 2 reduces sunitinib resistance in clear cell renal cell carcinoma.

doi: 10.4149/neo_2021_210823N1206

Figure Lengend Snippet: Figure 1. Expression pattern of STC2 in KIRC specimens and ccRCC cell lines. A) GEPIA analysis shows the mRNA expression levels of STC2 in 523 KIRCs and 100 non-KIRCs. B, C) The relative expression levels of STC2 in HK-2 cells and ccRCC cell lines were evalu ated by RT-qPCR and western blotting. β-actin was used as a loading reference. D) The STC2 contents in the culture medium of HK-2 cells and ccRCC cell lines were detected by ELISA. *p<0.05, **p<0.01

Article Snippet: Human STC2 neutralization antibody (AF2830), recombinant human STC2 (O76061), and STC2 ELISA Kit (CSB-EL022822HU) were purchased from R&D system (USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Neoplasma

Article Title: Blocking stanniocalcin 2 reduces sunitinib resistance in clear cell renal cell carcinoma.

doi: 10.4149/neo_2021_210823N1206

Figure Lengend Snippet: Figure 3. Effect of STC2 on sunitinib resistance in ccRCC cells. A–C) Determi nation of cell viability of Caki-1, 786-O, and 769-P cells in the absence (Ctrl) or presence of 5 μM sunitinib or 5 μM sunitinib+0.2 μg/ml STC2 neutralizing an tibody. D–F) Determination of cell viability of Caki-1, 786-O, and 769-P cells in the absence (Ctrl) or presence of 5 μM sunitinib or 5 μM sunitinib+500 ng/ ml hSTC2. *p<0.05

Article Snippet: Human STC2 neutralization antibody (AF2830), recombinant human STC2 (O76061), and STC2 ELISA Kit (CSB-EL022822HU) were purchased from R&D system (USA).

Techniques:

Journal: Neoplasma

Article Title: Blocking stanniocalcin 2 reduces sunitinib resistance in clear cell renal cell carcinoma.

doi: 10.4149/neo_2021_210823N1206

Figure Lengend Snippet: Figure 6. Effects of the STC2 neutralizing antibody on sunitinib accumulation, lysosomal pH, cell proliferation, and apoptosis in Caki-1 cells. A) Phase-contrast microscopy showing accumulation of yellow granules in Caki-1 cells incubated in the absence (Ctrl) or presence of sunitinib (5 μM), 5 μM sunitinib+0.2 μg/ml STC2 neutralizing antibody for 24 h. B, C) AO staining indicates the lysosome pH in Caki-1 cells incubated in the absence (Ctrl) or presence of sunitinib (5 μM), 5 μM sunitinib+0.2 μg/ml STC2 neutralizing antibody for 24 h. Quantification of the ratio between the red and green signal of AO was performed by the software ImageJ. D) The fluorescence of the lysosomal probe (LysoTracker™ Green DND-26) in Caki-1 cells incubated in the absence (Ctrl) or presence of sunitinib (5 μM), 5 μM sunitinib+0.2 μg/ml STC2 neutralizing antibody for 24 h. E, F) The immuno fluorescence staining of cell proliferation marker Ki-67 and cell apoptosis detection by TUNEL staining in Caki-1 cells incubated in the absence (Ctrl) or presence of sunitinib (5 μM), 5 μM sunitinib+0.2 μg/ml STC2 neutralizing antibody for 24 h. Quantification of Ki-67 and TUNEL fluorescence was performed by the software ImageJ. NS: no significant difference between control with treatments. *p<0.05

Article Snippet: Human STC2 neutralization antibody (AF2830), recombinant human STC2 (O76061), and STC2 ELISA Kit (CSB-EL022822HU) were purchased from R&D system (USA).

Techniques: Microscopy, Incubation, Staining, Software, Fluorescence, Marker, TUNEL Assay, Control

Journal: Oncology reports

Article Title: Clinical significance of stanniocalcin 2 expression as a predictor of tumor progression in gastric cancer.

doi: 10.3892/or.2013.2775

Figure Lengend Snippet: Figure 1. RT-PCR analysis of STC2 mRNA expression in cell lines and in blood specimens from patients with gastric cancer and healthy volunteers. Horizontal bars indicate mean STC2 mRNA copy numbers. STC2, stan niocalcin 2.

Article Snippet: The sections were incubated at room temperature for 60 min with an anti-human STC2 polyclonal antibody (Proteintech Group, Inc., Chicago, IL, USA) diluted 1:200 in Dako antibody diluent with background reducing components (DakoCytomation).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Oncology reports

Article Title: Clinical significance of stanniocalcin 2 expression as a predictor of tumor progression in gastric cancer.

doi: 10.3892/or.2013.2775

Figure Lengend Snippet: Figure 2. RT-PCR analysis of STC2 mRNA expression in cell spiking study. Serially diluted MKN-74 tumor cells (104, 103, 102, 101 and 0) were mixed with normal PBLs. Bars indicate standard deviation (SD). STC2, stannio calcin 2; PBLs, peripheral blood lymphocytes.

Article Snippet: The sections were incubated at room temperature for 60 min with an anti-human STC2 polyclonal antibody (Proteintech Group, Inc., Chicago, IL, USA) diluted 1:200 in Dako antibody diluent with background reducing components (DakoCytomation).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Standard Deviation

Journal: Oncology reports

Article Title: Clinical significance of stanniocalcin 2 expression as a predictor of tumor progression in gastric cancer.

doi: 10.3892/or.2013.2775

Figure Lengend Snippet: Figure 3. Kaplan-Meier survival curves for patients with gastric cancer based on the status of STC2 expression. Patients with STC2-positive expression had a significantly poorer prognosis than those with STC2-negative expression (P=0.014). STC2, stanniocalcin 2.

Article Snippet: The sections were incubated at room temperature for 60 min with an anti-human STC2 polyclonal antibody (Proteintech Group, Inc., Chicago, IL, USA) diluted 1:200 in Dako antibody diluent with background reducing components (DakoCytomation).

Techniques: Expressing

Journal: Oncology reports

Article Title: Clinical significance of stanniocalcin 2 expression as a predictor of tumor progression in gastric cancer.

doi: 10.3892/or.2013.2775

Figure Lengend Snippet: Figure 4. Representative immunohistochemical staining of STC2 expression in primary gastric tumor specimens. (A) Tumor cells with negative expression of STC2. (B) Tumor cells with weak expression of STC2. (C) Tumor cells with strong expression of STC2. Scale bar, 100 µm. Original magnification, x400. STC2, stanniocalcin 2.

Article Snippet: The sections were incubated at room temperature for 60 min with an anti-human STC2 polyclonal antibody (Proteintech Group, Inc., Chicago, IL, USA) diluted 1:200 in Dako antibody diluent with background reducing components (DakoCytomation).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Animals : an Open Access Journal from MDPI

Article Title: CircRNA8220 Sponges MiR-8516 to Regulate Cell Viability and Milk Synthesis via Ras/MEK/ERK and PI3K/AKT/mTOR Pathways in Goat Mammary Epithelial Cells

doi: 10.3390/ani10081347

Figure Lengend Snippet: RT-qPCR primers used in this study.

Article Snippet: To explore miR-8516 whether target gene STC2 and circRNA8220, 260 bp sequence of STC2 3′UTR and 344 bp total sequence of circRNA8220 were cloned into the psiCHECK-2 vectors (Addgene, CA, USA), respectively.

Techniques: Sequencing

Journal: Animals : an Open Access Journal from MDPI

Article Title: CircRNA8220 Sponges MiR-8516 to Regulate Cell Viability and Milk Synthesis via Ras/MEK/ERK and PI3K/AKT/mTOR Pathways in Goat Mammary Epithelial Cells

doi: 10.3390/ani10081347

Figure Lengend Snippet: Transfection efficiency of pcDNA3.1-STC2 and pCD5-circRNA8220 vectors and STC2-siRNA, circRNA8220-siRNA and miR-8516 mimics/inhibitors. ( a , b ) mRNA levels of circRNA8220. ( c ) mRNA levels of miR-8516. ( d ) The mRNA levels of STC2. ( e ) Protein levels of STC2. ** p < 0.01, * p < 0.05.

Article Snippet: To explore miR-8516 whether target gene STC2 and circRNA8220, 260 bp sequence of STC2 3′UTR and 344 bp total sequence of circRNA8220 were cloned into the psiCHECK-2 vectors (Addgene, CA, USA), respectively.

Techniques: Transfection

Journal: Animals : an Open Access Journal from MDPI

Article Title: CircRNA8220 Sponges MiR-8516 to Regulate Cell Viability and Milk Synthesis via Ras/MEK/ERK and PI3K/AKT/mTOR Pathways in Goat Mammary Epithelial Cells

doi: 10.3390/ani10081347

Figure Lengend Snippet: Antibody used in this study.

Article Snippet: To explore miR-8516 whether target gene STC2 and circRNA8220, 260 bp sequence of STC2 3′UTR and 344 bp total sequence of circRNA8220 were cloned into the psiCHECK-2 vectors (Addgene, CA, USA), respectively.

Techniques:

Journal: Animals : an Open Access Journal from MDPI

Article Title: CircRNA8220 Sponges MiR-8516 to Regulate Cell Viability and Milk Synthesis via Ras/MEK/ERK and PI3K/AKT/mTOR Pathways in Goat Mammary Epithelial Cells

doi: 10.3390/ani10081347

Figure Lengend Snippet: MiR-8516 down-regulated the expression level of STC2 via the 3′UTR. ( a ) The seed sequence of miR-8516 could match with WT-STC2-3′UTR and could not match with Mut-STC2-3′UTR. ( b ) Luciferase reporter assay of 293T cells co-transfected with WT-STC2-3′UTR or Mut-STC2-3′UTR and miR-8516 mimic, NC (negative control), miR-8516 inhibitor or NCH (NC-inhibitor). ( c ) miR-8516 decreased STC2 protein level in GMECs. ( d ) miR-8516 decreased STC2 mRNA level in GMECs. ** p < 0.01, * p < 0.05.

Article Snippet: To explore miR-8516 whether target gene STC2 and circRNA8220, 260 bp sequence of STC2 3′UTR and 344 bp total sequence of circRNA8220 were cloned into the psiCHECK-2 vectors (Addgene, CA, USA), respectively.

Techniques: Expressing, Sequencing, Luciferase, Reporter Assay, Transfection, Negative Control

Journal: Animals : an Open Access Journal from MDPI

Article Title: CircRNA8220 Sponges MiR-8516 to Regulate Cell Viability and Milk Synthesis via Ras/MEK/ERK and PI3K/AKT/mTOR Pathways in Goat Mammary Epithelial Cells

doi: 10.3390/ani10081347

Figure Lengend Snippet: circRNA8220 increased the expression of STC2 in GMEC in vitro. ( a ) si-circRNA8220 reduced the mRNA expression of STC2. ( b ) circRNA8220 increased the mRNA expression of STC2. ( c ) si-circRNA8220 blocked the protein expression of STC2. ( d ) circRNA8220 raised the protein expression of STC2. ** p < 0.01.

Article Snippet: To explore miR-8516 whether target gene STC2 and circRNA8220, 260 bp sequence of STC2 3′UTR and 344 bp total sequence of circRNA8220 were cloned into the psiCHECK-2 vectors (Addgene, CA, USA), respectively.

Techniques: Expressing, In Vitro

Journal: Animals : an Open Access Journal from MDPI

Article Title: CircRNA8220 Sponges MiR-8516 to Regulate Cell Viability and Milk Synthesis via Ras/MEK/ERK and PI3K/AKT/mTOR Pathways in Goat Mammary Epithelial Cells

doi: 10.3390/ani10081347

Figure Lengend Snippet: STC2 inhibited apoptosis and promoted proliferation of GMEC in vitro. ( a , b ) Cell proliferation was assessed using the cell counting kit-8 (CCK-8) assay after transfection with pc3.1-STC2 or si-STC2. ( c , d ) Cell proliferation indices were assessed after treatment with Edu after transfection with pc3.1-STC2 or si-STC2. ( e , f ) Apoptosis analysis of GMEC was detected with flow cytometry method after transfection with pc3.1-STC2 or si-STC2. ( g , h ) Protein levels of Bcl-2, Bax, caspase 3 and caspase 9 in GMEC after transfection with pc3.1-STC2 or si-STC2. Protein levels were measured by WB (western blot) densitometry was normalised to the β-actin density from the same lane. Data were expressed as the means ± SEM. ** p < 0.01, * p < 0.05.

Article Snippet: To explore miR-8516 whether target gene STC2 and circRNA8220, 260 bp sequence of STC2 3′UTR and 344 bp total sequence of circRNA8220 were cloned into the psiCHECK-2 vectors (Addgene, CA, USA), respectively.

Techniques: In Vitro, Cell Counting, CCK-8 Assay, Transfection, Flow Cytometry, Western Blot

Journal: Animals : an Open Access Journal from MDPI

Article Title: CircRNA8220 Sponges MiR-8516 to Regulate Cell Viability and Milk Synthesis via Ras/MEK/ERK and PI3K/AKT/mTOR Pathways in Goat Mammary Epithelial Cells

doi: 10.3390/ani10081347

Figure Lengend Snippet: STC2 enhanced the synthesis of β-casein and triglycerides in GMEC. ( a , b , c , d ) Secretion of β-casein and triglycerides (TG) after transfection with pc3.1-STC2 or si-STC2 was measured by enzyme-linked immunosorbent assay kit. ** p < 0.01, * p < 0.05.

Article Snippet: To explore miR-8516 whether target gene STC2 and circRNA8220, 260 bp sequence of STC2 3′UTR and 344 bp total sequence of circRNA8220 were cloned into the psiCHECK-2 vectors (Addgene, CA, USA), respectively.

Techniques: Transfection, Enzyme-linked Immunosorbent Assay

Journal: Animals : an Open Access Journal from MDPI

Article Title: CircRNA8220 Sponges MiR-8516 to Regulate Cell Viability and Milk Synthesis via Ras/MEK/ERK and PI3K/AKT/mTOR Pathways in Goat Mammary Epithelial Cells

doi: 10.3390/ani10081347

Figure Lengend Snippet: STC2 activated the signaling pathways of Ras/MEK/ERK and PI3K/AKT/mTOR in GMEC. ( a , b ) Western blot analysis was performed to detect the protein expression of Ras, p-MEK, MEK, p-ERK44/42, ERK44/42, p-PI3K, PI3K, p-AKT, AKT, p-mTOR, mTOR, p-S6K, S6K and β-actin in GMEC after transfection with pc3.1-STC2 or si-STC2. Protein levels were measured by WB densitometry was normalised to the β-actin density from the same lane. Data were expressed as the means ± SEM. ** p < 0.01, * p < 0.05.

Article Snippet: To explore miR-8516 whether target gene STC2 and circRNA8220, 260 bp sequence of STC2 3′UTR and 344 bp total sequence of circRNA8220 were cloned into the psiCHECK-2 vectors (Addgene, CA, USA), respectively.

Techniques: Protein-Protein interactions, Western Blot, Expressing, Transfection