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: The Journal of Pathology: Clinical Research

Article Title: Stanniocalcin 2 expression is associated with a favourable outcome in male breast cancer

doi: 10.1002/cjp2.106

Figure Lengend Snippet: STC2 overexpression in MBC. Significantly higher expression of STC2 was seen in MBC ( n = 12) compared to FBC ( n = 10) (A), also implied by RT‐qPCR analysis of three male and three female cases (B) and in cultured primary fibroblasts derived from male ( n = 4) and female ( n = 3) BC (C). While the number of MBC cases in the Oncomine™ analysis is low ( n = 4, compared with 322 females), data mining showed higher expression of STC2 in MBC versus FBC (D). Data on graphs are displayed as mean ± SD, except (D) where data are displayed as median, 90th percentile and 10th percentile (minimum and maximum values also shown). M, male; F, female.

Article Snippet: Sections were blocked with 1:10 Casein solution (Vector Laboratories ® #SP‐5020, Peterborough, UK) in antibody diluent (Thermo Fisher Scientific #003218) to block non‐specific staining, then incubated overnight at 4 °C with STC2 antibody (manufacturer: Atlas antibodies, supplier: Cambridge Bioscience, Cambridge, UK, HPA045372) solution 1:400 in antibody diluent (isotype controls were diluted to the same final concentration).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Cell Culture, Derivative Assay

Journal: The Journal of Pathology: Clinical Research

Article Title: Stanniocalcin 2 expression is associated with a favourable outcome in male breast cancer

doi: 10.1002/cjp2.106

Figure Lengend Snippet: Representative images of STC2 staining in MBC TMAs. The top panel shows examples of the various staining intensities in individual TMA cores, with higher magnification areas shown in the yellow outlined inserts. Black outlined inserts indicate foci of plasma membrane staining. The graph below shows the % of TMA samples which were categorised into each ‘intensity’ group (where the majority of tumour cells had at least the given intensity). +, weak, ++, medium staining, +++, strong staining. Images scanned at x20 objective magnification.

Article Snippet: Sections were blocked with 1:10 Casein solution (Vector Laboratories ® #SP‐5020, Peterborough, UK) in antibody diluent (Thermo Fisher Scientific #003218) to block non‐specific staining, then incubated overnight at 4 °C with STC2 antibody (manufacturer: Atlas antibodies, supplier: Cambridge Bioscience, Cambridge, UK, HPA045372) solution 1:400 in antibody diluent (isotype controls were diluted to the same final concentration).

Techniques: Staining, Clinical Proteomics, Membrane

Journal: The Journal of Pathology: Clinical Research

Article Title: Stanniocalcin 2 expression is associated with a favourable outcome in male breast cancer

doi: 10.1002/cjp2.106

Figure Lengend Snippet: Kaplan–Meier survival curves showing impact of STC2 staining H‐score in tumour and stroma on patient prognosis. High STC2 H‐scores in tumour cells (A), stroma (C) and total (E) were associated with longer DFS ( p = 0.017, p = 0.03, p = 0.035, respectively), but had no significant impact on OS for tumour (B), stroma (D) or total (F) ( p = 0.069, p = 0.65, p = 0.23). Grey line, high STC2 H‐score; black line, low STC2 H‐score, Log‐rank test. Cases were dichotomised by STC2 H‐score: H‐score cut‐off point was 108.5 for tumour cells (DFS n = 28 low, n = 23 high; OS n = 76 low, n = 73 high); 28.4 for stroma (DFS n = 16 low, n = 35 high; OS n = 49 low, n = 100 high) and 90.5 for total staining (DFS n = 28 low, n = 23 high; OS n = 77 low, n = 72 high). HR, hazard ratio, followed by confidence intervals shown in brackets.

Article Snippet: Sections were blocked with 1:10 Casein solution (Vector Laboratories ® #SP‐5020, Peterborough, UK) in antibody diluent (Thermo Fisher Scientific #003218) to block non‐specific staining, then incubated overnight at 4 °C with STC2 antibody (manufacturer: Atlas antibodies, supplier: Cambridge Bioscience, Cambridge, UK, HPA045372) solution 1:400 in antibody diluent (isotype controls were diluted to the same final concentration).

Techniques: Staining

Journal: The Journal of Pathology: Clinical Research

Article Title: Stanniocalcin 2 expression is associated with a favourable outcome in male breast cancer

doi: 10.1002/cjp2.106

Figure Lengend Snippet: Multivariate analysis of STC2 expression in MBC

Article Snippet: Sections were blocked with 1:10 Casein solution (Vector Laboratories ® #SP‐5020, Peterborough, UK) in antibody diluent (Thermo Fisher Scientific #003218) to block non‐specific staining, then incubated overnight at 4 °C with STC2 antibody (manufacturer: Atlas antibodies, supplier: Cambridge Bioscience, Cambridge, UK, HPA045372) solution 1:400 in antibody diluent (isotype controls were diluted to the same final concentration).

Techniques: Expressing

Journal: Frontiers in Neuroscience

Article Title: Stanniocalcin2 , but Not Stanniocalcin1 , Responds to Hypoxia in a HIF1-Dependent Manner in the Retina

doi: 10.3389/fnins.2022.882559

Figure Lengend Snippet: Primers used for semiquantitative real-time PCR.

Article Snippet: STC2 , Rabbit , Acris antibodies, Herford, Germany , 10314-1-AP , 1:300.

Techniques:

Journal: Frontiers in Neuroscience

Article Title: Stanniocalcin2 , but Not Stanniocalcin1 , Responds to Hypoxia in a HIF1-Dependent Manner in the Retina

doi: 10.3389/fnins.2022.882559

Figure Lengend Snippet: Antibodies used for immunofluorescence on retinal sections.

Article Snippet: STC2 , Rabbit , Acris antibodies, Herford, Germany , 10314-1-AP , 1:300.

Techniques: Immunofluorescence

Journal: Frontiers in Neuroscience

Article Title: Stanniocalcin2 , but Not Stanniocalcin1 , Responds to Hypoxia in a HIF1-Dependent Manner in the Retina

doi: 10.3389/fnins.2022.882559

Figure Lengend Snippet: Gene expression of Stc1 and Stc2 in the retina during postnatal development, aging and degeneration. (A) Relative expression levels of Stc1 (green bars) and Stc2 (blue bars) in different tissues isolated from 11 weeks old wild type (C57BL/6) mice. Relative expression levels of Stc1 (B) and Stc2 (C) in the retina of wild type (C57BL/6) mice (solid line) and rd10 mice (dotted line) from PND10 to PND180. Expression was determined by semi-quantitative real-time PCR and normalized to Actb levels. Shown are mean values ± SD of N = 3. * P < 0.05. Two-way ANOVA with Sidak’s multiple comparison test.

Article Snippet: STC2 , Rabbit , Acris antibodies, Herford, Germany , 10314-1-AP , 1:300.

Techniques: Expressing, Isolation, Real-time Polymerase Chain Reaction

Journal: Frontiers in Neuroscience

Article Title: Stanniocalcin2 , but Not Stanniocalcin1 , Responds to Hypoxia in a HIF1-Dependent Manner in the Retina

doi: 10.3389/fnins.2022.882559

Figure Lengend Snippet: Upregulation of Stc2 gene expression in response to hypoxia. (A) Relative expressions of Stc1 (left panel) and Stc2 (right panel) in retinas of 6 weeks old wild type (Balb/c) mice exposed to hypoxia (6 h, 7% O 2 ). Tissue was collected immediately after hypoxia (Imm) or after 2, 4, and 16 h of reoxygenation following hypoxia. (B) Relative expression of Stc1 , Stc2 , Adm , and Epo in retinas of wild-type (129S6) mice exposed to normoxia (black bars) or hypoxia (tissue collected immediately after 6 h at 7% O 2 ) (colored bars) at 7 weeks or 7 months of age. Expression levels were determined by semi-quantitative real-time PCR, normalized to Actb and shown relative to the levels in untreated controls (set to 1) (A) or to normoxic controls at 7 weeks of age (set to 1) (B) . Shown are mean values ± SD of N = 3. Significance was tested using one-way ANOVA with Dunnett’s multiple comparison test (A) and by Student’s t-test (B) (* P < 0.05, ** P < 0.01, *** P < 0.001, P < 0.0001****).

Article Snippet: STC2 , Rabbit , Acris antibodies, Herford, Germany , 10314-1-AP , 1:300.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in Neuroscience

Article Title: Stanniocalcin2 , but Not Stanniocalcin1 , Responds to Hypoxia in a HIF1-Dependent Manner in the Retina

doi: 10.3389/fnins.2022.882559

Figure Lengend Snippet: Site-specific detection of STC2 protein and mRNA expression in retinal tissue. (A) Immunostaining for STC2 (red) in a retinal flatmount of a 7 weeks old wild type (129S6) mouse. (B) Immunostaining for STC2 in a retinal section of an 11 weeks old wild type (129S6) mouse. Co-immunostainings for STC2 (red) and SYN ( C , green), GFAP ( D , green), GS ( E , green), and BRN3A ( F , green). Nuclei are stained with DAPI (blue). Scale bar: 50 μM. (G) Relative expression of Stc2 mRNA in the RPE (retinal pigment epithelium), ONL (outer nuclear layer), INL (inner nuclear layer), and GCL (ganglion cell layer) that were isolated by laser capture microdissection. (H) The amplified PCR products from experiments shown in panel (G) were run on an agarose gel for visualization. N = 3. (I) Relative expression of layer-specific control genes: Mct3 for RPE, Gnat1 for ONL, Vsx2 for INL, and Opn4 for GCL. Expression was determined by semi-quantitative real-time PCR and normalized to Actb . Shown are mean values ± SD of N = 3.

Article Snippet: STC2 , Rabbit , Acris antibodies, Herford, Germany , 10314-1-AP , 1:300.

Techniques: Expressing, Immunostaining, Staining, Isolation, Laser Capture Microdissection, Amplification, Agarose Gel Electrophoresis, Real-time Polymerase Chain Reaction

Journal: Frontiers in Neuroscience

Article Title: Stanniocalcin2 , but Not Stanniocalcin1 , Responds to Hypoxia in a HIF1-Dependent Manner in the Retina

doi: 10.3389/fnins.2022.882559

Figure Lengend Snippet: Upregulation of Stc2 upon stabilization of HIF1A in rod photoreceptors. Relative expression of Stc1 (green bars) and Stc2 (blue bars) in rod Δ Vhl , rod Δ Vhl;Hif 1 a , rod ΔVhl;Hif 2 a , and rod Δ Vhl;Hif 1 a;Hif 2 a mice and their control lines that carried the respective floxed genes without Opsin-cre (–). Expression was determined by semi-quantitative real-time PCR and normalized to Actb levels. Shown are mean values ± SD of N = 3. **** P < 0.0001. Two-way ANOVA with Sidak’s multiple comparison test.

Article Snippet: STC2 , Rabbit , Acris antibodies, Herford, Germany , 10314-1-AP , 1:300.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in Neuroscience

Article Title: Stanniocalcin2 , but Not Stanniocalcin1 , Responds to Hypoxia in a HIF1-Dependent Manner in the Retina

doi: 10.3389/fnins.2022.882559

Figure Lengend Snippet: Retinal morphology in the absence of STC1 and STC2. Retinal morphology of wild type (WT, C57BL/6) (A) , Stc1 –/– (B) and Stc2 –/– , (C) mice. (D) Spidergram of the retinal thickness of WT (C57BL/6, black line) and Stc2 –/– (blue line) mice; N = 3, ONH: Optic Nerve Head Immunostainings for STC2 (red) in retinal sections of wild type (C57BL/6) (E) , Stc1 –/– (F) , and Stc2 –/– (G) mice. Immunostainings for BRN3A (H) , SYN (I) , and GFAP (J) in retinal sections of Stc2 –/– mice. DAPI (blue) was used as nuclear stain. Images were acquired at 200 μm from the optic nerve head. Mice were 6 months (wild type) or 9 months ( Stc1 –/– and Stc2 –/– ) of age. Shown are representative images of N = 3 retinas per genotype. Scale bar: 50 μm.

Article Snippet: STC2 , Rabbit , Acris antibodies, Herford, Germany , 10314-1-AP , 1:300.

Techniques: Staining