Journal: Brazilian Journal of Physical Therapy

Article Title: Can transcutaneous electrical nerve stimulation improve achilles tendon healing in rats?

doi: 10.1590/bjpt-rbf.2014.0107

Figure Lengend Snippet: Comparison of the quantity of collagen types III (%) in the tendon tissue between the TENS and Simulation groups. Data expressed in mean and standard deviation.

Article Snippet: The expression of collagen I and III, observed in the immunohistochemical reaction with the polyclonal antibodies for the collagen types I and III (Affinity Purified Anti-Collagen Type I and Type III, Rabbit Anti-Human, Rockland Inc., Gilbertsville, PA, USA), was analyzed by quantifying the area corresponding to each type of collagen.

Techniques: Comparison, Standard Deviation

Journal: Brazilian Journal of Physical Therapy

Article Title: Can transcutaneous electrical nerve stimulation improve achilles tendon healing in rats?

doi: 10.1590/bjpt-rbf.2014.0107

Figure Lengend Snippet: Measures of variables related to group, assessment, and results of Analysis of Variance (ANOVA).

Article Snippet: The expression of collagen I and III, observed in the immunohistochemical reaction with the polyclonal antibodies for the collagen types I and III (Affinity Purified Anti-Collagen Type I and Type III, Rabbit Anti-Human, Rockland Inc., Gilbertsville, PA, USA), was analyzed by quantifying the area corresponding to each type of collagen.

Techniques: Standard Deviation

Journal: Scientific reports

Article Title: Prolactin-induced protein (PIP) increases the sensitivity of breast cancer cells to drug-induced apoptosis.

doi: 10.1038/s41598-023-33707-w

Figure Lengend Snippet: Figure 1. Characteristics of human breast cancer (BC) cell lines with overexpression or suppressed expression of PIP. (A) Expression of PIP mRNA in parental (wild type) MDA-MB-231 cells (MDA-231.WT), control MDA-MB-231 transduced with the vector alone (MDA-231.C), MDA-MB-231 cells overexpressing PIP (MDA- 231.PIP), and parental (wild type) T47D cells (T47D.WT), control T47D transduced with the vector alone (T47D.shC), and T47D cells with suppressed expression of the PIP gene (T47D.shPIP). The relative level of PIP mRNA expression was determined by real-time PCR. PIP expression levels were normalized against the SDHA gene and MCF-7 cells served as the calibrator sample. The results are expressed as mean ± SD. (B) Western blotting analysis of anti-PIP rabbit monoclonal antibody binding to proteins present in lysates and cultures media of MDA-231.WT, MDA-231.C, MDA-231.PIP cells and T47D.WT, T47D.shC, T47D.shPIP. Cell lysates (equivalent to 25 μg of protein) and culture supernatants (equivalent to 25 μg of protein) were separated by SDS- PAGE under reducing conditions on 15% gel and electrophoretically transferred onto nitrocellulose membrane. For cell lysates, GAPDH was served as an internal control. Viability of BC MDA-231.PIP cells overexpressing PIP and T47D.shPIP cells with suppressed expression of PIP grown in the presence of increasing concentrations of anti-cancer drugs: doxorubicin (DOX) (C, D), 4-hydroperoxycyclophosphamide (4-HC) (E, F) and paclitaxel (PAX) (G and H) for 48 h. The percentage of viable cells was determined using the MTT assay as described in the “Materials and methods”. (C) Cells grown in the absence of anti-cancer drugs. Data represent the mean ± SD of four replicates from three independent measurements. Statistically significant differences (*p < 0.1, **p < 0.01, ***p < 0.001).

Article Snippet: To detect PIP and apoptotic proteins such as CRADD, DAPK1 and CD40, the following antibodies were used: rabbit monoclonal anti-PIP (clone EP1582Y, 1:1000, Abcam, UK), rabbit monoclonal anti-CRADD (clone ARC1771, 1:1000, St. JohnsLaboratory, UK), rabbit polyclonal anti-DAPK1 (1:1000, Cell Signaling, MA, USA) and rabbit monoclonal anti-CD40 (clone D8W3N, 1:1000, Cell Signaling, MA, USA).

Techniques: Over Expression, Expressing, Control, Transduction, Plasmid Preparation, Real-time Polymerase Chain Reaction, Western Blot, Binding Assay, SDS Page, Membrane, MTT Assay

Journal: Scientific reports

Article Title: Prolactin-induced protein (PIP) increases the sensitivity of breast cancer cells to drug-induced apoptosis.

doi: 10.1038/s41598-023-33707-w

Figure Lengend Snippet: Figure 3. Fold expression changes of up-regulated pro-apoptotic genes. (A) MDA-231.PIP vs. MDA-231.C and down-regulated pro-apoptotic genes in T47D-shPIP vs. T47.shC cells quantified by qPCR. The bar graphs represent the fold regulation of mRNAs as the mean ± standard deviation (n = 2). (B) The qPCR data were validated by Western blotting analysis of protein lysates from MDA-231.PIP, MDA-231.C, T47D.shPIP, and T47D.shC cells using specific antibodies.

Article Snippet: To detect PIP and apoptotic proteins such as CRADD, DAPK1 and CD40, the following antibodies were used: rabbit monoclonal anti-PIP (clone EP1582Y, 1:1000, Abcam, UK), rabbit monoclonal anti-CRADD (clone ARC1771, 1:1000, St. JohnsLaboratory, UK), rabbit polyclonal anti-DAPK1 (1:1000, Cell Signaling, MA, USA) and rabbit monoclonal anti-CD40 (clone D8W3N, 1:1000, Cell Signaling, MA, USA).

Techniques: Expressing, Standard Deviation, Western Blot

Journal: Scientific reports

Article Title: Prolactin-induced protein (PIP) increases the sensitivity of breast cancer cells to drug-induced apoptosis.

doi: 10.1038/s41598-023-33707-w

Figure Lengend Snippet: Figure 4. (A) Xenograft tumor growth of MDA-231.PIP with overexpression of PIP and control MDA-231.C; T47D.shPIP with the suppressed expression of PIP and control T47D.shC cells in nude Crl:NU(Ncr) mice. (B) Mice bearing tumors were subjected to treatment with DOX. Impact of DOX on the growth of (C) MDA-231. PIP, MDA-231.C cells, (D) T47D.shPIP and T47D.shC cells. MDA-231.PIP/placebo and MDA-231.C/placebo— mice with MDA-231.PIP or MDA-231.C tumors treated with placebo. MDA-231.PIP/DOX and MDA-231.C/ DOX—mice with MDA-231.PIP or MDA-231.C tumors treated with DOX; T47D.shPIP/placebo or T47D.shC/ placebo mice with T47D.shPIP or T47D.shC tumors treated with placebo. T47D.shPIP/DOX and T47D.shC/ DOX—mice with T47D.shPIP or T47D.shC tumors treated with DOX. Data were shown as the mean tumor volume for the group of mice (n = 10–12 for MDA-231 cells and n = 7–8 for T47D cells) ± SD at each indicated time point (arrows). Data were analyzed using Graphpad prism 7.0 two-way Anova Dunnett’s post test. (E) The number of apoptotic cells and (F) Ki-67-positive cells in MDA-231.PIP, MDA-231.C, T47D.shPIP, and T47D.shC tumors determined by TUNEL assay and IHC staining with monoclonal antibody against Ki-67 and compared by the Mann–Whitney U test (**p < 0.01, ***p < 0.001).

Article Snippet: To detect PIP and apoptotic proteins such as CRADD, DAPK1 and CD40, the following antibodies were used: rabbit monoclonal anti-PIP (clone EP1582Y, 1:1000, Abcam, UK), rabbit monoclonal anti-CRADD (clone ARC1771, 1:1000, St. JohnsLaboratory, UK), rabbit polyclonal anti-DAPK1 (1:1000, Cell Signaling, MA, USA) and rabbit monoclonal anti-CD40 (clone D8W3N, 1:1000, Cell Signaling, MA, USA).

Techniques: Over Expression, Control, Expressing, TUNEL Assay, Immunohistochemistry, MANN-WHITNEY

Journal: Scientific reports

Article Title: Prolactin-induced protein (PIP) increases the sensitivity of breast cancer cells to drug-induced apoptosis.

doi: 10.1038/s41598-023-33707-w

Figure Lengend Snippet: Figure 6. (A) Coomassie-blue-stained SDS-PAGE and (B) immunostaining with anti-PIP rabbit monoclonal antibody of recombinant PIP (PIPlexsy). Lines 1–4 represent PIPlexsy fractions purified by affinity chromatography on NiNTA resin using elution buffer containing 500 mM, 200 mM, 100 mM, and 50 mM imidazole, respectively. 1–5 µg protein was subjected to SDS-PAGE under reducing conditions in 15% gel. (C) Flow cytometric analysis of PIPlexsy binding to BC MDA-MB-231 cells. (1) Flow cytometry dot plots show the percentage of PIP-positive cells detected with monoclonal rabbit antibody against PIP and secondary goat antibodies directed against rabbit IgG conjugated with Alexa Fluor488. (2) Binding of monoclonal rabbit antibody against PIP and secondary goat antibodies directed against rabbit IgG conjugated with Alexa Fluor488 in the absence of PIPlexsy. (D) PIP increases the sensitivity of MDA-MB-231 cells to DOX-induced apoptosis. MDA-MB-231 cells were incubated with 0.1 or 10 µg of PIPlexsy in the absence or presence of 0.5 µM DOX. C—control MDA-MB-231 cells grown in the absence of PIPlexsy and DOX.

Article Snippet: To detect PIP and apoptotic proteins such as CRADD, DAPK1 and CD40, the following antibodies were used: rabbit monoclonal anti-PIP (clone EP1582Y, 1:1000, Abcam, UK), rabbit monoclonal anti-CRADD (clone ARC1771, 1:1000, St. JohnsLaboratory, UK), rabbit polyclonal anti-DAPK1 (1:1000, Cell Signaling, MA, USA) and rabbit monoclonal anti-CD40 (clone D8W3N, 1:1000, Cell Signaling, MA, USA).

Techniques: Staining, SDS Page, Immunostaining, Recombinant, Purification, Affinity Chromatography, Binding Assay, Flow Cytometry, Incubation, Control

Journal: AMB Express

Article Title: Quantitative proteomics revealed modulation of macrophages by MetQ gene of Streptococcus suis serotype 2

doi: 10.1186/s13568-020-01131-2

Figure Lengend Snippet: Validation of proteomics results. a Seven genes encoding for the corresponding differentially expressed proteins were selected to verify the proteomics results by qPCR. The results were calculated by 2 −△△CT method. b Pstpip1, Src and Hmox1 proteins were selected for the validation of the quantitative proteomics. GAPDH was used as reference. The gray intensities are shown on the right. ZY05719, ZY05719 treated cells; △ MetQ , △ MetQ treated cells; control, non-infected cells. The statistical significance was determined by Student’s t test. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Membranes were incubated with primary antibodies Src (Proteintech), Pstpip1 (Proteintech), Hmox1 (Abcam) and GAPDH (CMCTAG) overnight at 4 °C.

Techniques: Biomarker Discovery, Quantitative Proteomics, Control, Infection

Journal: AMB Express

Article Title: Quantitative proteomics revealed modulation of macrophages by MetQ gene of Streptococcus suis serotype 2

doi: 10.1186/s13568-020-01131-2

Figure Lengend Snippet: Immune system process related proteins through GO annotation

Article Snippet: Membranes were incubated with primary antibodies Src (Proteintech), Pstpip1 (Proteintech), Hmox1 (Abcam) and GAPDH (CMCTAG) overnight at 4 °C.

Techniques: Cell Differentiation

Journal: Pathogens

Article Title: Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

doi: 10.3390/pathogens9080611

Figure Lengend Snippet: Binding of fluorescent receptor binding proteins (RBP) fusion reporters of Yersinia phages L-413C and ΦA1122 to Y. pestis cells. ( a ) mCherry fused to RBP of phage L-413C (red signals) or ( b ) eGFP fused to RBP of phage ΦA1122 (green signals) were added to cells of logarithmically growing Y. pestis cultures, washed with buffer to remove unbound reporter proteins, transferred to microscopy slides and subjected to fluorescence microscopy. Shown are representative fluorescence signals recorded at 630× magnification (scale bar: 5 µm).

Article Snippet: Taken together, the proposed function of GpH (of phage L-413C) and Gp17 (of phage ΦA1122) by employing fluorescent fusion protein technology could be experimentally supported.

Techniques: Binding Assay, Microscopy, Fluorescence

Journal: Pathogens

Article Title: Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

doi: 10.3390/pathogens9080611

Figure Lengend Snippet: Time course of binding of Yersinia phage RBP fusion reporters to Y. pestis cells. Cultures of Y. pestis EV76 were grown from lag to exponential (logarithmic) and stationary phase and samples withdrawn at representative time points for RBP binding assays. Individual micrographs are shown for mCherry-RBP fusion protein (phage L-413C; red signal) or eGFP-RBP fusion protein (phage ΦA1122; green signal) binding to Y. pestis cells at 6, 20, 28 and 37 °C (recorded in merged light and fluorescence channels; scale bar: 5 µm).

Article Snippet: Taken together, the proposed function of GpH (of phage L-413C) and Gp17 (of phage ΦA1122) by employing fluorescent fusion protein technology could be experimentally supported.

Techniques: Binding Assay, Fluorescence

Journal: Pathogens

Article Title: Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

doi: 10.3390/pathogens9080611

Figure Lengend Snippet: Semiquantitative representation of binding of Yersinia phage RBP fusion reporters to Y. pestis cells. Micrographs taken from culture samples withdrawn at different time points (growth phases) from growth experiments at 6, 20, 28 or 37 °C (see for representative examples) were visually analyzed for RBP-reporter binding to Y. pestis EV76 cells. ( a ) mCherry-RBP (phage L-413C; red) or ( b ) eGFP-RBP (phage ΦA1122; green) fusion proteins. Binding of the reporters was assessed by an increase or decrease (light colors) of fluorescence signals (and phases with constant strong signals; dark colors) and plotted onto data (black dots) from actual growth curve experiments (measurements of optical culture densities at 600 nm). Periods with lack of visual fluorescence signals indicating no RBP binding, were scored as grey lines.

Article Snippet: Taken together, the proposed function of GpH (of phage L-413C) and Gp17 (of phage ΦA1122) by employing fluorescent fusion protein technology could be experimentally supported.

Techniques: Binding Assay, Fluorescence

Journal: Pathogens

Article Title: Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

doi: 10.3390/pathogens9080611

Figure Lengend Snippet: Binding of RBP fusion reporters to growing cultures of Y. pestis EV76 cells at 37 °C. In this representation, capsule formation is detected by means of monoclonal anti-F1 capsule antigen antibody in combination with secondary antibody labelled with Alexa Fluor 488 (green fluorescence, upper panels only) for co-detection with L-413C-RBP or Alexa Fluor 647 (false color blue fluorescence, lower panels only) for co-detection with ΦA1122-RPB, respectively. RBP binding to Y. pestis EV76 cells at indicated time points at 37 °C is shown as individual representative micrographs for phage L-413C mCherry-RBP-reporter (red signals) or phage ΦA1122 eGFP-RBP-reporter (green signals) as merges with fluorescent antibody signals (and recorded in merged light and fluorescence channels). Arrows indicate doubly labelled cells with RBP-reporters and antibodies (scale bar: 5 µm).

Article Snippet: Taken together, the proposed function of GpH (of phage L-413C) and Gp17 (of phage ΦA1122) by employing fluorescent fusion protein technology could be experimentally supported.

Techniques: Binding Assay, Fluorescence

Journal: Pathogens

Article Title: Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

doi: 10.3390/pathogens9080611

Figure Lengend Snippet: Specificity tests of the RBP-reporters binding to cells of Yersinia spp. closely related to Y. pestis . Cultures of several Yersinia spp. (as indicated) and of Y. pestis EV76 as positive control were grown at 28 °C ( a ), or 37 °C ( b ). After 4 h of growth, samples were withdrawn, incubated with mCherry-RBP-reporter (from phage L-413C; red signals) or eGFP-RBP-reporter (from phage ΦA1122; green signals) and subjected to fluorescence microscopy (recorded in merged light and fluorescence channels). The arrow indicates an individual cell labelled by the RBP-reporter (scale bar: 5 µm).

Article Snippet: Taken together, the proposed function of GpH (of phage L-413C) and Gp17 (of phage ΦA1122) by employing fluorescent fusion protein technology could be experimentally supported.

Techniques: Binding Assay, Positive Control, Incubation, Fluorescence, Microscopy

Journal: Pathogens

Article Title: Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

doi: 10.3390/pathogens9080611

Figure Lengend Snippet: Binding of RBP-reporters to cells of additional Y. pseudotuberculosis strains at 37 °C. Cultures and samples of three Y. pseudotuberculosis O:1a strains were treated as described in ), mCherry-RBP (from phage L-413C; red signals) or eGFP-RBP fusion proteins (from phage ΦA1122; green signals) were added and samples subjected to fluorescence microscopy (recorded in merged light and fluorescence channels; scale bar: 5 µm).

Article Snippet: Taken together, the proposed function of GpH (of phage L-413C) and Gp17 (of phage ΦA1122) by employing fluorescent fusion protein technology could be experimentally supported.

Techniques: Binding Assay, Fluorescence, Microscopy

Journal: Pathogens

Article Title: Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

doi: 10.3390/pathogens9080611

Figure Lengend Snippet: Binding of RBP-reporters to inactivated cells of Y. pestis . Cells of Y. pestis EV76 grown for 7 h at 28 °C were inactivated as indicated, washed with PBS buffer, incubated with fusion proteins mCherry-RBP (phage L-413C; red signals, upper panels) or mCherry-RBP (phage ΦA1122; red signals lower panels) and subjected to fluorescence microscopy (recorded in merged light and fluorescence channels). Untreated cells served as positive controls (scale bar: 5 µm).

Article Snippet: Taken together, the proposed function of GpH (of phage L-413C) and Gp17 (of phage ΦA1122) by employing fluorescent fusion protein technology could be experimentally supported.

Techniques: Binding Assay, Incubation, Fluorescence, Microscopy

Journal: Pathogens

Article Title: Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

doi: 10.3390/pathogens9080611

Figure Lengend Snippet: Binding of RBP-reporters to inactivated cells of RG-3 Y. pestis . Cells of Y. pestis grown for 7 h at 28 °C on solid BHI media were inactivated with 4% PFA. Inactivated cells were incubated with fusion proteins mCherry-RBP (phage L-413C; red signals, upper panels) or eGFP-RBP (phage ΦA1122; green signals lower panels) and subjected to fluorescence microscopy (recorded in merged light and fluorescence channels). Strains tested represent a phylogenetically diverse set of Y. pestis isolates as indicated (scale bar: 5 µm).

Article Snippet: Taken together, the proposed function of GpH (of phage L-413C) and Gp17 (of phage ΦA1122) by employing fluorescent fusion protein technology could be experimentally supported.

Techniques: Binding Assay, Incubation, Fluorescence, Microscopy