Journal: International Journal of Molecular Sciences

Article Title: The Transcription Factor SCX is a Potential Serum Biomarker of Fibrotic Diseases

doi: 10.3390/ijms21145012

Figure Lengend Snippet: SCX expression levels in lung fibroblasts correlated with the expression of profibrotic genes. Expression of genes SCX ( a ), COL1A1 ( b ), COL1A2 ( c ), and α-SMA/ACTA2 ( d ) in pulmonary fibroblasts CCD25Lu and CCD8Lu. (a) Scleraxis expression was measured by qPCR using Taqman Hs03054634_g1 relative to GAPDH (Hs02786624_g1). Panels (b–d) correspond to COL1A1 (Hs00164004_m1), COL1A2 (Hs01028956_m1), and α-SMA/ACTA2 (Hs00426835_g1) expression relative to GAPDH, respectively. Statistical differences were assessed using the Student’s t-test. ** = p < 0.01, *** = p < 0.001.

Article Snippet: Real-time PCR was performed using LightCycler ® 480 System (Roche, Basel, Switzerland) with TaqMan probes (Thermo Scientific, Waltham, MA, USA) labeled with FAM (Hs03054634_g1 for SCX, Hs00164004_m1 for COL1A1, Hs01028970_m1 for COL1A2, Hs00426835_g1 for α-SMA/ACTA2, and Hs01012685_m1 for TCF3) and VIC (Hs02786624_g1 for GAPDH and Hs0280069S_m1 for HPRT).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Transcription Factor SCX is a Potential Serum Biomarker of Fibrotic Diseases

doi: 10.3390/ijms21145012

Figure Lengend Snippet: Gene expression of SCX and its target genes was high in IPF fibroblast primary cultures. mRNA levels of the indicated genes were quantified as in using cDNA samples from primary lung fibroblast cultures from a healthy donor (NOVA) and three IPF patients (HIPF). Relative expression of SCX ( a ), COL1A1 ( b ), COL1A2 ( c ), α-SMA/ACTA2 ( d ), and TCF3 ( e ). The statistical test used for this assay was one-way ANOVA. *** = p < 0.001.

Article Snippet: Real-time PCR was performed using LightCycler ® 480 System (Roche, Basel, Switzerland) with TaqMan probes (Thermo Scientific, Waltham, MA, USA) labeled with FAM (Hs03054634_g1 for SCX, Hs00164004_m1 for COL1A1, Hs01028970_m1 for COL1A2, Hs00426835_g1 for α-SMA/ACTA2, and Hs01012685_m1 for TCF3) and VIC (Hs02786624_g1 for GAPDH and Hs0280069S_m1 for HPRT).

Techniques: Gene Expression, Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Transcription Factor SCX is a Potential Serum Biomarker of Fibrotic Diseases

doi: 10.3390/ijms21145012

Figure Lengend Snippet: SCX expression was elevated in IPF tissue. Pulmonary tissue derived from IPF patients showed higher expression of SCX in comparison to healthy controls. Immunohistochemical analyses were made in two tissue samples from healthy controls (samples 573-11 and 860-05) and three IPF patients (425-05, 428-16, and 442-16). Samples were stained with hematoxylin–eosin and Masson’s trichrome. SCX was localized with a rabbit polyclonal antibody against human SCX (1:25; Antibodies online ABIN9670006). The fibroblastic foci in all IPF samples were localized in the center of each picture. In the hematoxylin–eosin stain, the fibroblastic foci were observed as pale areas composed of elongated cells, whose nuclei appeared stained with SCX (in the SCX stain). In the Masson’s trichome stain, the fibroblastic foci also showed blue coloring due to the presence of collagen.

Article Snippet: Real-time PCR was performed using LightCycler ® 480 System (Roche, Basel, Switzerland) with TaqMan probes (Thermo Scientific, Waltham, MA, USA) labeled with FAM (Hs03054634_g1 for SCX, Hs00164004_m1 for COL1A1, Hs01028970_m1 for COL1A2, Hs00426835_g1 for α-SMA/ACTA2, and Hs01012685_m1 for TCF3) and VIC (Hs02786624_g1 for GAPDH and Hs0280069S_m1 for HPRT).

Techniques: Expressing, Derivative Assay, Comparison, Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: The Transcription Factor SCX is a Potential Serum Biomarker of Fibrotic Diseases

doi: 10.3390/ijms21145012

Figure Lengend Snippet: SCX serum concentrations were significantly higher in patients with IPF and SSc compared to controls. SCX serum levels in patients with HP were not different from controls but differed significantly from patients with IPF and SSc. *** = p < 0.001.

Article Snippet: Real-time PCR was performed using LightCycler ® 480 System (Roche, Basel, Switzerland) with TaqMan probes (Thermo Scientific, Waltham, MA, USA) labeled with FAM (Hs03054634_g1 for SCX, Hs00164004_m1 for COL1A1, Hs01028970_m1 for COL1A2, Hs00426835_g1 for α-SMA/ACTA2, and Hs01012685_m1 for TCF3) and VIC (Hs02786624_g1 for GAPDH and Hs0280069S_m1 for HPRT).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Transcription Factor SCX is a Potential Serum Biomarker of Fibrotic Diseases

doi: 10.3390/ijms21145012

Figure Lengend Snippet: IPF patients with severely affected pulmonary function showed higher levels of circulating SCX. SCX serum levels grouped by ( a ) smoking habit and hypertension, ( b ) BMI, ( c ) sex and age, ( d ) TLC and FVC, and ( e ) DLCO. includes an excel file with raw data. Higher SCX serum levels were found in patients with severely diminished TLC (≤ 60%) and with FVC below 80%. Ex-Smoker: patient that has not smoked for at least one year. Non-Smoker: patient with no smoke habit. Hypertension: values above 139 mmHg in systolic pressure and 89 mmHg in diastolic pressure were considered as hypertension . * = p < 0.05, ** = p < 0.01. Total Lung Capacity (TLC): maximum volume of air present in the lungs. A normal person can inhale approximately 6 L. Forced Vital Capacity (FVC): volume of air that can be forcibly blown out after a full inspiration. Normal values are approximately 4.6 L .

Article Snippet: Real-time PCR was performed using LightCycler ® 480 System (Roche, Basel, Switzerland) with TaqMan probes (Thermo Scientific, Waltham, MA, USA) labeled with FAM (Hs03054634_g1 for SCX, Hs00164004_m1 for COL1A1, Hs01028970_m1 for COL1A2, Hs00426835_g1 for α-SMA/ACTA2, and Hs01012685_m1 for TCF3) and VIC (Hs02786624_g1 for GAPDH and Hs0280069S_m1 for HPRT).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Transcription Factor SCX is a Potential Serum Biomarker of Fibrotic Diseases

doi: 10.3390/ijms21145012

Figure Lengend Snippet: SCX serum levels were high in late progression SSc patients and patients suffering associated pulmonary fibrosis. SCX circulating levels were grouped by ( a ) smoking habit and BMI, ( b ) serologic profile, ( c ) sex and age, ( d ) internal organ involvement, ( e ) classification and progression, and ( f ) FVC. includes an excel file with raw data. Secondary organ involvement comprised pulmonary fibrosis (PF), scleroderma renal crisis (SCR), and gastrointestinal involvement (GI). The GI included esophageal dysmotility, esophageal stricture, small bowel hypomotility or dilation, small bowel bacterial overgrowth, malabsorption syndrome, or use of parenteral nutrition. (d) SSc classification and progression. * = p < 0.05. ** = p < 0.01. *** = p < 0.001. Scl70 = anti-topoisomerase I antibodies.

Article Snippet: Real-time PCR was performed using LightCycler ® 480 System (Roche, Basel, Switzerland) with TaqMan probes (Thermo Scientific, Waltham, MA, USA) labeled with FAM (Hs03054634_g1 for SCX, Hs00164004_m1 for COL1A1, Hs01028970_m1 for COL1A2, Hs00426835_g1 for α-SMA/ACTA2, and Hs01012685_m1 for TCF3) and VIC (Hs02786624_g1 for GAPDH and Hs0280069S_m1 for HPRT).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Transcription Factor SCX is a Potential Serum Biomarker of Fibrotic Diseases

doi: 10.3390/ijms21145012

Figure Lengend Snippet: SCX overexpression promoted an induction in α-SMA expression that was dependent on TCF3. ( a ) The expression of the indicated proteins was analyzed using immunoblots. CCD8Lu cells were transduced with adenoviral vectors for 48 h. The infective particles included an empty vector as a control, and adenoviral vectors encoding SCX, TCF3, and a combination of SCX and TCF3. ( b ) Densitometric analyses of α-SMA induction relative to β-actin, quantifying three independent samples of each transduction. A one-way ANOVA test confirmed statistical differences in α-SMA expression in empty vs. TCF3-SCX samples. * p = 0.0086.

Article Snippet: Real-time PCR was performed using LightCycler ® 480 System (Roche, Basel, Switzerland) with TaqMan probes (Thermo Scientific, Waltham, MA, USA) labeled with FAM (Hs03054634_g1 for SCX, Hs00164004_m1 for COL1A1, Hs01028970_m1 for COL1A2, Hs00426835_g1 for α-SMA/ACTA2, and Hs01012685_m1 for TCF3) and VIC (Hs02786624_g1 for GAPDH and Hs0280069S_m1 for HPRT).

Techniques: Over Expression, Expressing, Western Blot, Transduction, Plasmid Preparation, Control

Journal: Scientific reports

Article Title: Polymeric human Fc-fusion proteins with modified effector functions.

doi: 10.1038/srep00124

Figure Lengend Snippet: Figure 6 | Binding of polymeric Fc-fusions to CD191 human B-lymphocytes. (a) Characteristic FACs plot showing different populations of human leucocytes represented by their forward (FSC) and side (SSC) profiles. (b) Individual CD191 B-lymphocytes stained with anti-human CD19-FITC (shown by blue dots) were gated. (c) Binding of 40 mg of hexameric (blue trace) and dimeric (green trace) PfMSP119-hIgG1-Fc-LH309/310CL-TP to the gated human CD191 B lymphocytes was detected using phycoerythrin (PE)-labeled goat (Fab92) anti-hIgG. No binding was detected with the phycoerythrin (PE)-labeled goat (Fab92) anti-hIgG in the absence of Fc-fusion proteins (red trace). Data are representative of three separate experiments.

Article Snippet: Cells were washed twice with FACs buffer and incubated for 1h at 4uC with 1/500 dilution of F(ab9)2 goat anti-hIgG-Fc-phycoerythrin (PE) or goat anti-hCD19-fluorescein isothiocyanate (FITC)-conjugated Abs (Southern Biotechnology) in 200 ml FACs buffer.

Techniques: Binding Assay, Staining, Labeling