Journal: PLoS ONE

Article Title: The Human Homolog of Drosophila Headcase Acts as a Tumor Suppressor through Its Blocking Effect on the Cell Cycle in Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0137579

Figure Lengend Snippet: Association between the HECA homo protein quantity and clinicopathologic features of 93 patients with HCC.

Article Snippet: The primary antibodies included mouse polyclonal antibody against human HECA (Abnova, Taiwan, 1:1000) and rabbit polyclonal antibody against β-actin (1:1000, Sigma-Aldrich, USA).

Techniques:

Journal: PLoS ONE

Article Title: The Human Homolog of Drosophila Headcase Acts as a Tumor Suppressor through Its Blocking Effect on the Cell Cycle in Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0137579

Figure Lengend Snippet: (A) The HECA homo mRNA level was the highest in the HepG2 cell lines among the five examined cell lines. (B) For each of the three cell lines (HepG2, Huh-7, and MHCC-97H), the HECA homo mRNA level in the siRNA group (cells transfected with specific siRNA against HECA homo) was significantly lower than in the SNC group (cells that were transfected with scrambled siRNA as a negative control) or in the blank group (cells treated with only Lipofectamine 2000). In contrast, there was no significant difference between the SNC group and the blank group. (C) The HECA homo mRNA level in the HECA exp group (cells that were transfected with HECA homo full-length-expressing plasmid) was significantly higher than those in the mock group (cells that were transfected with empty control plasmid) or blank group (cells that were treated with only Lipofectamine 2000). Meanwhile, there was no significant difference between the mock group and the blank group. (D) and (E) the corresponding protein levels were detected by western blotting.

Article Snippet: The primary antibodies included mouse polyclonal antibody against human HECA (Abnova, Taiwan, 1:1000) and rabbit polyclonal antibody against β-actin (1:1000, Sigma-Aldrich, USA).

Techniques: Transfection, Negative Control, Expressing, Plasmid Preparation, Control, Western Blot

Journal: PLoS ONE

Article Title: The Human Homolog of Drosophila Headcase Acts as a Tumor Suppressor through Its Blocking Effect on the Cell Cycle in Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0137579

Figure Lengend Snippet: (A) Cell proliferation viability as assessed by MTT. For each of the three cell lines, the siRNA group exhibited more viability than did the SNC group, whereas the HECA exp group exhibited less viability than did the mock group. (B) Representative pictures for the cell colony formation assay (right) and quantitative analysis of colony number (left). For each of the three cell lines, more colonies were found in the siRNA group than in the SNC group, whereas fewer colonies were found in the HECA exp group than in the mock group.

Article Snippet: The primary antibodies included mouse polyclonal antibody against human HECA (Abnova, Taiwan, 1:1000) and rabbit polyclonal antibody against β-actin (1:1000, Sigma-Aldrich, USA).

Techniques: Colony Assay

Journal: PLoS ONE

Article Title: The Human Homolog of Drosophila Headcase Acts as a Tumor Suppressor through Its Blocking Effect on the Cell Cycle in Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0137579

Figure Lengend Snippet: (A) Representative pictures of cell cycle distribution as detected by flow cytometric analysis (left). Comparison of the relative cell numbers in the G1 phase between groups (right). For each of the three cell lines, fewer cells in the G1 phase were detected in the siRNA group than in the SNC group. And more cells in the G1 phase were found in the HECA exp group than in the mock group. (B) Representative pictures of apoptosis as detected by flow cytometric analysis (left). Comparison of the apoptosis rate (proportion of cells in early and late apoptotic stages) between groups (right). For each of the three cell lines, the apoptosis rate of the siRNA group was not significantly different from that of the SNC group, For Huh-7 and MHCC-97H cells, the apoptosis rate of the HECA exp group was also not significantly different from that of the mock group. But for HepG2 cells, the apoptosis rate of the HECA exp group was significantly higher than that of the mock group.

Article Snippet: The primary antibodies included mouse polyclonal antibody against human HECA (Abnova, Taiwan, 1:1000) and rabbit polyclonal antibody against β-actin (1:1000, Sigma-Aldrich, USA).

Techniques: Comparison

Journal: The Journal of Neuroscience

Article Title: Localization of PDZD7 to the Stereocilia Ankle-Link Associates this Scaffolding Protein with the Usher Syndrome Protein Network

doi: 10.1523/JNEUROSCI.3071-12.2012

Figure Lengend Snippet: Detection of PDZD7 in hair cells stereocilia. A–D, Mass spectrometry analysis. A, C, Representative MS2 spectra for PDZD7 and GPR98 showing fragments that were either matched (colored) or unmatched (black) to hypothetical database spectra. The peptide sequence and statistical score indicating identification confidence are shown for each. B, D, Total peptide coverage of PDZD7 and GPR98. The width of each bar indicates the length of the identified peptide, the position of the bar along the x-axis corresponds to its position in the sequence, the height of the bar indicates the number of identical peptides identified, and the shading of the bar corresponds to log(e), the statistical significance for peptide identification, averaged over all identical peptides. The statistical score indicating identification confidence is shown for each protein. E–N, PDZD7 immunolocalization in developing rat hair cells. Confocal microscopy of inner (IHC; E, H, I, J and N), outer (OHC; F, M) and vestibular (VHC; G, K, L) hair cells from postnatal (P) day 2 (E–I), 7 (J–L) and 10 (M, N) rats immunostained with mouse polyclonal YF-PA20973 (mAb) and rabbit polyclonal PB960 antibodies, showing localization of PDZD7 (red) at the stereocilia ankle-link region; actin (green). H, High magnification of a long and a short stereocilium from a P2 inner hair cell revealing the PDZD7 fluorescence just above the tapered base. J, Horizontal cross-section through IHC stereocilia showing that PDZD7 immunofluorescence (red) is peripheral to the actin core (green); it is abundant between stereocilia of the same row and between stereocilia of longer and shorter rows, but excluded from the back of the tallest stereocilia. K, Oblique view of P7 vestibular stereocilia showing PDZD7 fluorescence (red, inset) at the ankle-link region. L, Absence of PDZD7 (red) at the upper tip-link insertion sites where myosin VIIa (blue) is clustered (arrowheads). O–Q, Maturing hair cells expressing mCherry-PDZD7 showing that PDZD7 targets the ankle-link region. (Scale bars: 5 μm, except in H: 300 nm)

Article Snippet: A mouse polyclonal anti-PDZD7 antibody (YF-PA20973, called mAb in figure legends) directed against human PDZD7 amino-acids 1–511 ( {"type":"entrez-protein","attrs":{"text":"AAH29054","term_id":"20810507","term_text":"AAH29054"}} AAH29054 ) was purchased from AbFrontier (Seoul, Korea).

Techniques: Mass Spectrometry, Sequencing, Confocal Microscopy, Fluorescence, Immunofluorescence, Expressing

Journal: The Journal of Neuroscience

Article Title: Localization of PDZD7 to the Stereocilia Ankle-Link Associates this Scaffolding Protein with the Usher Syndrome Protein Network

doi: 10.1523/JNEUROSCI.3071-12.2012

Figure Lengend Snippet: PDZD7 colocalization with ankle-link proteins in maturing rat hair cells. A–C, Colocalization of PDZD7 and taperin in hair cells. A, B, Back (A) and orthogonal (B) views of stereocilia rows of P10 OHCs co-labeled for PDZD7 (blue) and taperin (green) showing the distribution of both PDZD7 (blue) and taperin (green) along the long and short stereocilia; actin (red). C, A cartoon summarizing PDZD7 (blue) and taperin (green) distribution along the stereocilia (red). D–H, PDZD7 and whirlin colocalization in developing hair cells. Whirlin is revealed with antibodies GP66 (D–F) and GP68 (G, H). D, Oblique view of P8 OHC hair bundle showing immunofluorescence distribution of PDZD7 (P, green) and whirlin (W, red); actin (blue). E, H, Oblique view of P8 VHC stereocilia showing immunofluorescence distribution of PDZD7 (green) and whirlin (blue); actin (red). F, A view of a P2 vestibular hair bundle showing partial colocalization of PDZD7 (green) and whirlin (blue); R=0.64 and Rr=0.60; actin (red). G, Orthogonal view through P2 OHC hair bundles showing PDZD7 (green) and whirlin (blue) immunofluorescence at the ankle-link region; actin (red). I–K, PDZD7 and GPR98 colocalization in P4 hair cells. I, Back view of P4 OHC showing PDZD7 (green) and GPR98 (blue) colocalization at the ankle-link region. J, K, Oblique (K) and surface views (L) of P4 VHCs showing PDZD7 (green) and GPR98 (blue) colocalization at the ankle-link region (R=0.73; Rr=0.81 in K). L–N, Whirlin and GPR98 colocalization in P4 hair cells. L, A view of P4 OHC showing whirlin (green) and GPR98 (blue) colocalization at the ankle-link region. M, A view of P4 IHC showing whirlin (green) and GPR98 (blue) colocalization at the ankle-link region. N, Front lateral view of P4 VHC showing whirlin (green) and GPR98 (blue) presence at the ankle-link region. In all three hair cell types, whirlin is also seen at the stereocilia tips (L–N). (Scale bars: 5 μm in A–H, J, K, N; 2.5 μm in I, L, M)

Article Snippet: A mouse polyclonal anti-PDZD7 antibody (YF-PA20973, called mAb in figure legends) directed against human PDZD7 amino-acids 1–511 ( {"type":"entrez-protein","attrs":{"text":"AAH29054","term_id":"20810507","term_text":"AAH29054"}} AAH29054 ) was purchased from AbFrontier (Seoul, Korea).

Techniques: Labeling, Immunofluorescence

Journal: The Journal of Neuroscience

Article Title: Localization of PDZD7 to the Stereocilia Ankle-Link Associates this Scaffolding Protein with the Usher Syndrome Protein Network

doi: 10.1523/JNEUROSCI.3071-12.2012

Figure Lengend Snippet: PDZD7 colocalization with stereocilia ankle-link proteins in mature rat hair cells. A–D, PDZD7 (P) and GPR98 (G) colocalization in P15 hair cells. Both GPR98 (green) and PDZD7 (blue) are absent from mature IHCs (A) and OHCs (B), following the disappearance of ankle-links; actin (red). In mature VHCs where ankle-links persist, GPR98 (green) and PDZD7 (blue) fluorescence strongly overlap, as seen through longitudinal and oblique views of the stereocilia. E–I, PDZD7 (P) and whirlin (W) colocalization in P15 hair cells. In organ of Corti, PDZD7 (blue) is absent from mature IHCs (E, F) and OHCs (G) following the disappearance of ankle-links. Whirlin fluorescence (green) persists in stereocilia tips and some accumulation is visible around the ankle-link region (arrowhead in F). Longitudinal (H) and oblique (I) views of mature VHCs shows that PDZD7 fluorescence (blue) covers the regions where whirlin fluorescence (green) is present around the ankle-link area. In mature VHCs, whirlin is also seen concentrated at the stereocilia tips (H, I). J, Scanning electron micrograph of P10 whirler IHC stereocilia showing short stereocilia that are inter-connected with all sorts of links, including the ankle-links (arrowheads in K). (Scale bars: 5 μm, except in K: 1μm)

Article Snippet: A mouse polyclonal anti-PDZD7 antibody (YF-PA20973, called mAb in figure legends) directed against human PDZD7 amino-acids 1–511 ( {"type":"entrez-protein","attrs":{"text":"AAH29054","term_id":"20810507","term_text":"AAH29054"}} AAH29054 ) was purchased from AbFrontier (Seoul, Korea).

Techniques: Fluorescence

Journal: The Journal of Neuroscience

Article Title: Localization of PDZD7 to the Stereocilia Ankle-Link Associates this Scaffolding Protein with the Usher Syndrome Protein Network

doi: 10.1523/JNEUROSCI.3071-12.2012

Figure Lengend Snippet: A–F, PDZD7 interactions with Usher proteins in COS7 and LLC-PK1 cells. A, In the presence of EGFP-sans (green), mCherry-PDZD7 (red) is randomly spread in the COS7 cell cytosol, associated with actin filaments (blue), or also recruited to EGFP-sans clusters, as shown by the high values of Mander’s (R=0.80) and Pearson’s (Rr=0.81) coefficients. B, EGFP-harmonin-b (green) forms plaques associated with actin (blue). The distribution of mCherry-PDZD7 (red) is unchanged in the presence of EGFP-harmonin-b, and its colocalization with EGFP-harmonin-b actin-bound plaques is non-significant based on the values of R=0.80 and Rr=0.10. C, F, Confocal cross-section scans of the apical plasma membrane of polarized LLC-PK1 cells co-expressing fusion proteins IL2α-ush2a (green in C and red in E) and IL2α-GPR98 (green in D and red in F) together with either mCherry-PDZD7 (red in C, D) or EGFP-Whirlin (green in E, F), showing that both fusion proteins are enriched in the apical plasma membrane where they highly colocalize with both whirlin and PDZD7. (Scale bars: 10 μm)

Article Snippet: A mouse polyclonal anti-PDZD7 antibody (YF-PA20973, called mAb in figure legends) directed against human PDZD7 amino-acids 1–511 ( {"type":"entrez-protein","attrs":{"text":"AAH29054","term_id":"20810507","term_text":"AAH29054"}} AAH29054 ) was purchased from AbFrontier (Seoul, Korea).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Chimeric Proteins Containing MAP-1 and Functional Domains of C4b-Binding Protein Reveal Strong Complement Inhibitory Capacities

doi: 10.3389/fimmu.2018.01945

Figure Lengend Snippet: Recombinant constructs and purified proteins. (A) Diagram of the domain distribution of MAP-1, C4BP α-chain and the recombinant proteins. (B) Instant blue stain of purified MAP-1:C4BP 1−5 , C4BP 1−5 :MAP-1, and C4BP 1−5 via affinity chromatography. (C) The identity of the protein bands was confirmed by Western blotting using anti-MAP-1 mAb and rMAP-1 as positive control.

Article Snippet: The membranes were probed with 0.5 μg/ml anti-MAP-1 20C4 or 0.3 μg/ml anti-human C4BP polyclonal antibody (pAb; Abnova, 722-D01P, Taiwan) overnight, followed by 1 h incubation with 0.4 μg/ml rabbit anti-mouse-HRP pAb (Dako, P0260, Denmark) or a 1:4000 dilution of donkey anti-rabbit-HRP pAb (GE Healthcare, NA934V) for MAP-1 and C4BP, respectively.

Techniques: Recombinant, Construct, Purification, Staining, Affinity Chromatography, Western Blot, Positive Control

Journal: Frontiers in Immunology

Article Title: Chimeric Proteins Containing MAP-1 and Functional Domains of C4b-Binding Protein Reveal Strong Complement Inhibitory Capacities

doi: 10.3389/fimmu.2018.01945

Figure Lengend Snippet: Gel filtration chromatography of chimeric proteins. (A) Gel filtration profile of MAP-1:C4BP 1−5 (M:C) and C4BP 1−5 :MAP-1 (C:M) under physiological calcium conditions or 10 mM EDTA. The identity of the gel filtration profiles was confirmed by analyzing the elution fractions of M:C (B) or C:M (C) by sandwich ELISA using anti-C4BP as capture antibody and anti-MASP-1/-3/MAP-1 as detection. Relative abundance of the chimeric proteins in the elution fractions following gel filtration under calcium conditions or EDTA is expressed as OD. mAU, milli absorption units.

Article Snippet: The membranes were probed with 0.5 μg/ml anti-MAP-1 20C4 or 0.3 μg/ml anti-human C4BP polyclonal antibody (pAb; Abnova, 722-D01P, Taiwan) overnight, followed by 1 h incubation with 0.4 μg/ml rabbit anti-mouse-HRP pAb (Dako, P0260, Denmark) or a 1:4000 dilution of donkey anti-rabbit-HRP pAb (GE Healthcare, NA934V) for MAP-1 and C4BP, respectively.

Techniques: Filtration, Chromatography, Sandwich ELISA

Journal: Frontiers in Immunology

Article Title: Chimeric Proteins Containing MAP-1 and Functional Domains of C4b-Binding Protein Reveal Strong Complement Inhibitory Capacities

doi: 10.3389/fimmu.2018.01945

Figure Lengend Snippet: MAP-1:C4BP 1−5 (M:C), C4BP 1−5 :MAP-1 (C:M) and MAP-1 binding to MBL (A) and CL-11 (B) . rMBL and rCL-11 were immobilized onto mannan-coated plates. Recombinant proteins were applied in a two-fold dilution in the presence of calcium or EDTA. Binding was determined using an anti-MAP-1 mAb. Connecting lines display nonlinear fitting using the equation specific binding with hill slope. Results are representative of three independent experiments and error bars represent minimum and maximum values of triplicate measurements.

Article Snippet: The membranes were probed with 0.5 μg/ml anti-MAP-1 20C4 or 0.3 μg/ml anti-human C4BP polyclonal antibody (pAb; Abnova, 722-D01P, Taiwan) overnight, followed by 1 h incubation with 0.4 μg/ml rabbit anti-mouse-HRP pAb (Dako, P0260, Denmark) or a 1:4000 dilution of donkey anti-rabbit-HRP pAb (GE Healthcare, NA934V) for MAP-1 and C4BP, respectively.

Techniques: Binding Assay, Recombinant

Journal: Frontiers in Immunology

Article Title: Chimeric Proteins Containing MAP-1 and Functional Domains of C4b-Binding Protein Reveal Strong Complement Inhibitory Capacities

doi: 10.3389/fimmu.2018.01945

Figure Lengend Snippet: Chimeric and rMAP-1 complement inhibition after pre-incubation on rMBL/mannan. Serial dilutions of MAP-1:C4BP 1−5 (M:C), C4BP 1−5 :MAP-1 (C:M), and rMAP-1 were allowed to form complexes with mannan-bound rMBL prior to addition of MBL-defect serum as a source of complement. Detection of C4 (A) , C3 (B) , and TCC (C) deposition was quantified using anti-C4, anti-C3, and anti-TCC mouse mAbs. Connecting lines are four parameters nonlinear fitting using the equation inhibitor concentration vs. slope. Data are reported as [(OD inhibitor -OD background )/(OD noinhibitor -OD background )]100. Error bars represent the SEM of three independent experiments, and the dashed line the 50% inhibition level. * P < 0.05; *** P < 0.001; **** P < 0.0001.

Article Snippet: The membranes were probed with 0.5 μg/ml anti-MAP-1 20C4 or 0.3 μg/ml anti-human C4BP polyclonal antibody (pAb; Abnova, 722-D01P, Taiwan) overnight, followed by 1 h incubation with 0.4 μg/ml rabbit anti-mouse-HRP pAb (Dako, P0260, Denmark) or a 1:4000 dilution of donkey anti-rabbit-HRP pAb (GE Healthcare, NA934V) for MAP-1 and C4BP, respectively.

Techniques: Inhibition, Incubation, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Chimeric Proteins Containing MAP-1 and Functional Domains of C4b-Binding Protein Reveal Strong Complement Inhibitory Capacities

doi: 10.3389/fimmu.2018.01945

Figure Lengend Snippet: Chimeric and rMAP-1 complement inhibition after co-incubation with NHS. Serial dilutions of MAP-1:C4BP 1−5 (M:C), C4BP 1−5 :MAP-1 (C:M), and rMAP-1 were co-incubated with 2% NHS in non-adsorbent titration plates for 30 min prior to addition to mannan-bound rMBL. Detection of deposition of C4 (A) , C3 (B) , and TCC (C) was determined as described before. Connecting lines are four parameters nonlinear fitting using the equation inhibitor concentration vs. slope. Dashed line Data are reported as [(OD inhibitor – OD background )/(OD noinhibitor – OD background )]100. Error bars represent the SEM of three independent experiments, and the dashed line the 50% inhibition level. * P < 0.05; *** P < 0.001.

Article Snippet: The membranes were probed with 0.5 μg/ml anti-MAP-1 20C4 or 0.3 μg/ml anti-human C4BP polyclonal antibody (pAb; Abnova, 722-D01P, Taiwan) overnight, followed by 1 h incubation with 0.4 μg/ml rabbit anti-mouse-HRP pAb (Dako, P0260, Denmark) or a 1:4000 dilution of donkey anti-rabbit-HRP pAb (GE Healthcare, NA934V) for MAP-1 and C4BP, respectively.

Techniques: Inhibition, Incubation, Titration, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Chimeric Proteins Containing MAP-1 and Functional Domains of C4b-Binding Protein Reveal Strong Complement Inhibitory Capacities

doi: 10.3389/fimmu.2018.01945

Figure Lengend Snippet: Cofactor activity in factor I-mediated C4b and C3b cleavage. Purified C3b or C4b was incubated with fI and different cofactors for 2 h at 37°C. The reactions were stopped with LDS buffer and subjected to western blotting under reducing conditions using C4d and C3d antibodies. (A) Incubation of C4b with fI in the presence of C4BP, fH or C4BP-containing chimeric proteins resulted in the generation of a band corresponding to the inactive degradation products iC4b and C4d. (B) Incubation of C3b with fI in the presence of C4BP, fH or C4BP-containing chimeric proteins results in the generation of the 68 kDa degradation fragment. M:C, MAP-1:C4BP 1−5 ; C:M, C4BP 1−5 :MAP-1. The blots are representative of three independent experiments.

Article Snippet: The membranes were probed with 0.5 μg/ml anti-MAP-1 20C4 or 0.3 μg/ml anti-human C4BP polyclonal antibody (pAb; Abnova, 722-D01P, Taiwan) overnight, followed by 1 h incubation with 0.4 μg/ml rabbit anti-mouse-HRP pAb (Dako, P0260, Denmark) or a 1:4000 dilution of donkey anti-rabbit-HRP pAb (GE Healthcare, NA934V) for MAP-1 and C4BP, respectively.

Techniques: Activity Assay, Purification, Incubation, Western Blot

Journal: Frontiers in Immunology

Article Title: Chimeric Proteins Containing MAP-1 and Functional Domains of C4b-Binding Protein Reveal Strong Complement Inhibitory Capacities

doi: 10.3389/fimmu.2018.01945

Figure Lengend Snippet: Inhibition of complement deposition on tubular proximal kidney epithelial cells. (A) HK-2 cells were incubated with increasing concentrations of rMBL at 4°C and bound MBL was determined using an anti-MBL mAb. (B) Binding of MBL results in the deposition of C4. HK-2 cells were incubated with rMBL followed by 10% MBL defect serum for 1 h at 4°C. Bound C4 was determined using an anti-C4c pAb. (C) rMBL-mediated C4 deposition after MAP-1:C4BP 1−5 (M:C), C4BP 1−5 :MAP-1 (C:M), rMAP-1 or full length C4BP co-incubation with 10% MBL defect serum. Inhibition is reported as the ratio of the MFI of the inhibitor to the MFI of the bank (MFI inh :MFI blank ). Significance was tested for each concentration of the inhibitors compared to the blank. Data are reported as the mean ± SEM of three independent experiments.

Article Snippet: The membranes were probed with 0.5 μg/ml anti-MAP-1 20C4 or 0.3 μg/ml anti-human C4BP polyclonal antibody (pAb; Abnova, 722-D01P, Taiwan) overnight, followed by 1 h incubation with 0.4 μg/ml rabbit anti-mouse-HRP pAb (Dako, P0260, Denmark) or a 1:4000 dilution of donkey anti-rabbit-HRP pAb (GE Healthcare, NA934V) for MAP-1 and C4BP, respectively.

Techniques: Inhibition, Incubation, Binding Assay, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Chimeric Proteins Containing MAP-1 and Functional Domains of C4b-Binding Protein Reveal Strong Complement Inhibitory Capacities

doi: 10.3389/fimmu.2018.01945

Figure Lengend Snippet: Wielisa total complement screen. Serial dilutions of MAP-1:C4BP 1−5 (M:C), C4BP 1−5 :MAP-1 (C:M), rMAP-1, rC4BP 1−5 , and rMAP-1 and rC4BP 1−5 together (M+C) were incubated with NHS and subsequently the protein/serum mix was applied to pre-coated Wielisa plates. Complement activation was quantified using an anti-TCC mAb. (A) Lectin pathway activation on mannan-coated plates. (B) Classical pathway activation on IgM-coated plates. (C) Alternative activation on LPS-coated plates. Connecting lines are nonlinear fitting using the equation inhibitor concentration vs. slope. Values represent OD readings normalized to the OD of the wells without inhibitor times 100. Significance was tested for best-fit IC50 values of data sets with an adjusted R-squared > 0.8. Error bars represent the SEM of three independent experiments, and the dashed line the 50% inhibition level. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: The membranes were probed with 0.5 μg/ml anti-MAP-1 20C4 or 0.3 μg/ml anti-human C4BP polyclonal antibody (pAb; Abnova, 722-D01P, Taiwan) overnight, followed by 1 h incubation with 0.4 μg/ml rabbit anti-mouse-HRP pAb (Dako, P0260, Denmark) or a 1:4000 dilution of donkey anti-rabbit-HRP pAb (GE Healthcare, NA934V) for MAP-1 and C4BP, respectively.

Techniques: Incubation, Activation Assay, Concentration Assay, Inhibition