Journal: Journal of Neuroscience

Article Title: Kisspeptin Directly Excites Anorexigenic Proopiomelanocortin Neurons but Inhibits Orexigenic Neuropeptide Y Cells by an Indirect Synaptic Mechanism

doi: 10.1523/jneurosci.2098-10.2010

Figure Lengend Snippet: Figure 5. Kisspeptin induces an inward current in POMC cells. The inward current was recorded at a holding potential of 60 mV with TTX (0.5 M) in the bath. A, Kisspeptin (1 M) evoked an inward current in a POMC neuron in normal ACSF. B, The inward current induced by kisspeptin (1 M) in nominal zero-Ca 2/10 mM Mg 2 ACSF solution. C, The kisspeptin induced inward current in a POMC neuron was blocked with BAPTA (10 mM) in the pipette solution. D, Inward current induced by kisspeptin on a POMC cell in an ACSF solution with Na replaced by Li . E, The kisspeptin induced current on a POMC cell in the presence of NCX blocker KB-R7943 (60 M). F, Kisspeptin induced current in a POMC cell in the presence of a more selective NCX blocker SN-6 (10 M). G, Kisspeptin (1 M) inducedasmallinwardcurrentinthepresenceofSKF96365(10M).H,Bargraphshowingthemean

Article Snippet: Tetrodotoxin (TTX), 1-[2-(4-methoxyphenyl)-2-[3-(4-methoxyphenyl)propoxy]ethyl-1 Himidazole hydrochloride (SKF96365), 2-[2-[4-(4-nitrobenzyloxy) phenyl]ethyl]isothiourea mesylate (KB-R7943), 2-[[4-[(4-nitrophenyl) methoxy]phenyl]methyl]-4-thiazolidinecarboxylic acid ethyl ester (SN-6), and (S)-3,5-dihydroxyphenylglycine (DHPG) were obtained from Tocris Bioscience.

Techniques: Transferring

Journal: Journal of Neuroscience

Article Title: Kisspeptin Directly Excites Anorexigenic Proopiomelanocortin Neurons but Inhibits Orexigenic Neuropeptide Y Cells by an Indirect Synaptic Mechanism

doi: 10.1523/jneurosci.2098-10.2010

Figure Lengend Snippet: Figure 6. Kisspeptin-evoked current is attenuated by Li and KB-R7943. The current traces were evoked by voltage ramps from 130to0mVataholdingpotentialof 60mV.TTX(0.5M),AP5(50M),CNQX(10M),BIC(30M),andCd 2(200M) wereaddedinthebathsolution.A,RecordingsperformedwithanormalACSFbathperfusionandaKMeSO4pipettesolution.Top, TracesshowingthecurrentresponsestothevoltagerampinPOMCcellsbefore(Cont)andduringkisspeptin(Kiss)(1M).Bottom, The Kiss-induced current (subtraction of Cont from Kiss). B, Cont and Kiss (top) and Kiss-induced current (bottom) recorded with BaCl2 (0.3 mM) in the bath perfusion and a CsMeSO4 pipette solution. C, Kiss-induced current recorded using a CsMeSO4 pipette solution in modified ACSF with Na replaced by Li . D, Kiss-induced current recorded using a CsMeSO4 pipette solution after pretreatmentwithNCXblockerKB-R7943(60M).E,BargraphshowingtheamplitudeofKiss-inducedcurrentat 125mVunder different recording conditions. *p 0.05; **p 0.01 vs normal ACSF; group t test. Error bars indicate SEM.

Article Snippet: Tetrodotoxin (TTX), 1-[2-(4-methoxyphenyl)-2-[3-(4-methoxyphenyl)propoxy]ethyl-1 Himidazole hydrochloride (SKF96365), 2-[2-[4-(4-nitrobenzyloxy) phenyl]ethyl]isothiourea mesylate (KB-R7943), 2-[[4-[(4-nitrophenyl) methoxy]phenyl]methyl]-4-thiazolidinecarboxylic acid ethyl ester (SN-6), and (S)-3,5-dihydroxyphenylglycine (DHPG) were obtained from Tocris Bioscience.

Techniques: Transferring, Modification

Journal: Cell communication and signaling : CCS

Article Title: T-type voltage-gated channels, Na + /Ca 2+ -exchanger, and calpain-2 promote photoreceptor cell death in inherited retinal degeneration.

doi: 10.1186/s12964-023-01391-y

Figure Lengend Snippet: Fig. 6 Differential correlation of calpain-1 and calpain-2 to photoreceptor cell death. A Spearman analysis comparing the numbers of activated calpain-1 and calpain-2 in the outer nuclear layer (ONL) of untreated rd1 retina and rd1 retina treated with the calpain-2 inhibitor NA-184. B Spearman analysis between the numbers of activated calpain-1 and dying, TUNEL positive cells in untreated or NA-184 treated rd1 ONL. C Spearman analysis between the ratio of calpain-2/calpain-1 and TUNEL positive cells in untreated or NA-184 treated rd1 ONL. D Alluvial diagram showing the relationship between activated calpain-2 and calpain-1, as well as TUNEL positive cells in rd1 ONL after treatment with BAPTA-AM, L-cis, CM4620, SN-6, D-cis, TTA-AS, DS5565, and NA-184

Article Snippet: After explantation, the complete R16 medium was changed every two days along with the pharmaceutical compounds including 10 μM BAPTA-AM [44] (ab120503; Abcam, Cambridge, UK), 50μM L-cis-diltiazem [45] (ab120532; Abcam), 20 μM CM4620 [46] (HY-101942; MedChemExpress, Sollentuna, Sweden), 40 μM SN-6 [47] (HY-107658; MedChemExpress), 100 μM D-cis-diltiazem [45] (ab120260; Abcam), 10 μM TTA-A2 [48] (HY-111828; MedChemExpress), 15 μM DS5565 [49] (HY-108006; MedChemExpress), and 1 μM NA-184 [50] (kindly provided by Michel Baudry, Western University, Pomona, CA, USA), respectively.

Techniques: TUNEL Assay

Journal: Scientific Reports

Article Title: CD105 (Endoglin) as negative prognostic factor in AML

doi: 10.1038/s41598-019-54767-x

Figure Lengend Snippet: Generation and functional characterization of CD105 antibody K-ro23. The CD105 antibody K-ro23 was generated as described in the methods section. ( A ) Size exclusion chromatography after the initial purification with protein A affinity chromatography. ( B ) SDS-PAGE analysis under reducing (R) and non-reducing conditions (NR). ( C ) Flow cytometry based detection of specific binding of the CD105 antibody (shaded peaks) and an isotype control (open peaks) at 1 nM on CD105 transfected Sp2/0 and mock transfected Sp2/0, and ( D ) analysis of the binding capacity of the CD105 antibody on CD105 transfected and mock transfected Sp2/0 as negative control. ( E ) Comparison of the specific binding of K-ro23 and the commercial clones SN6 and 166707 (shaded peaks) compared to the respective isotype controls (open peaks) at 5 nM on acute leukaemia cell lines. ( F ) Titration of the different CD105 clones on acute leukaemia cell lines. Specific fluorescence intensity (SFI) levels are shown.

Article Snippet: Unconjugated anti-CD105 mAb (clone K-ro23 and clone SN6, Santa Cruz Biotechnology, Dallas, TX) or isotype control (clone MPC-11, BD Biosciences, Heidelberg, Germany) as well as the comparative anti-CD105 clone 166707 (R&D Systems, Minneapolis, MN) and the respective isotype control (IgG1 isotype, clone MG1-45, BioLegend) were added at 5 nM, followed by a mouse-specific PE conjugate (1:100).

Techniques: Functional Assay, Generated, Size-exclusion Chromatography, Purification, Affinity Chromatography, SDS Page, Flow Cytometry, Binding Assay, Control, Transfection, Negative Control, Comparison, Clone Assay, Titration, Fluorescence

Journal: Scientific Reports

Article Title: CD105 (Endoglin) as negative prognostic factor in AML

doi: 10.1038/s41598-019-54767-x

Figure Lengend Snippet: CD105 expression on malignant haematopoietic cells. ( A ) Immunohistochemistry of CD105 expression of two exemplary AML patients. ( B ) Exemplary gating for two AML samples with the respective isotype control and specific CD105 binding. Specific gating strategy and controls are shown in Fig. S2. ( C – G ) CD105 expression was analysed on AML cells by flow cytometry. SFI levels above 1.5 were considered as positive expression (dotted line) ( C ) CD105 expression on blasts of AML patients (n = 62) are depicted as percentage of CD105 positive blasts and SFI levels (boxplots with Tukey whiskers). ( D – F ) CD105 SFI levels are depicted for the different FAB types (single values, median; Kruskal-Wallis-test) ( D ), FAB M3 vs. others FAB (boxplots with Tukey whiskers; Mann-Whitney-test) ( E ) and according to favourable and unfavourable FAB classification (boxplots with Tukey whiskers; Mann-Whitney-test) ( F ) are shown. ( G ) Distribution of CD105 expression (SFI) throughout NCCN risk group (boxplots with Tukey whiskers; Kruskal-Wallis-test).

Article Snippet: Unconjugated anti-CD105 mAb (clone K-ro23 and clone SN6, Santa Cruz Biotechnology, Dallas, TX) or isotype control (clone MPC-11, BD Biosciences, Heidelberg, Germany) as well as the comparative anti-CD105 clone 166707 (R&D Systems, Minneapolis, MN) and the respective isotype control (IgG1 isotype, clone MG1-45, BioLegend) were added at 5 nM, followed by a mouse-specific PE conjugate (1:100).

Techniques: Expressing, Immunohistochemistry, Control, Binding Assay, Flow Cytometry, MANN-WHITNEY

Journal: Scientific Reports

Article Title: CD105 (Endoglin) as negative prognostic factor in AML

doi: 10.1038/s41598-019-54767-x

Figure Lengend Snippet: Impact of CD105 expression on clinical outcome. ( A ) Specific binding of the CD105 antibody (shaded peaks) and an isotype control (open peaks) at 5 nM on AML blasts from exemplary patients of quartiles 1–4 using flow cytometry. ( B ) Kaplan-Meier analysis of SFI levels of CD105. Overall survival (OS) of each SFI quartile (1 st : 1–2.42; 2 nd : 2.43–5.25; 3 rd :5.26–26.37; 4 th : 26.38–65.2) was plotted followed by statistical analysis with log-rank test. ( C ) ROC for SFI levels of CD105 and OS were plotted. The crossing of the dotted line and the ROC curve marks the Youden-index for the highest sensitivity and specificity. ( D ) Overall survival according to CD105 lo and CD105 hi expression in Kaplan-Meier analysis. Mean OS was reached in CD105 hi after 199 days (dotted line; log-rank test). ( E ) PFS after any AML specific treatment according to CD105 lo and CD105 hi expression in Kaplan-Meier analysis. In CD105 hi the mean PFS was 383 days (dotted line; log-rank test).

Article Snippet: Unconjugated anti-CD105 mAb (clone K-ro23 and clone SN6, Santa Cruz Biotechnology, Dallas, TX) or isotype control (clone MPC-11, BD Biosciences, Heidelberg, Germany) as well as the comparative anti-CD105 clone 166707 (R&D Systems, Minneapolis, MN) and the respective isotype control (IgG1 isotype, clone MG1-45, BioLegend) were added at 5 nM, followed by a mouse-specific PE conjugate (1:100).

Techniques: Expressing, Binding Assay, Control, Flow Cytometry

Journal: Scientific Reports

Article Title: CD105 (Endoglin) as negative prognostic factor in AML

doi: 10.1038/s41598-019-54767-x

Figure Lengend Snippet: Distribution of patient characteristics according to CD105 quartiles.

Article Snippet: Unconjugated anti-CD105 mAb (clone K-ro23 and clone SN6, Santa Cruz Biotechnology, Dallas, TX) or isotype control (clone MPC-11, BD Biosciences, Heidelberg, Germany) as well as the comparative anti-CD105 clone 166707 (R&D Systems, Minneapolis, MN) and the respective isotype control (IgG1 isotype, clone MG1-45, BioLegend) were added at 5 nM, followed by a mouse-specific PE conjugate (1:100).

Techniques:

Journal: Scientific Reports

Article Title: CD105 (Endoglin) as negative prognostic factor in AML

doi: 10.1038/s41598-019-54767-x

Figure Lengend Snippet: Distribution of patient characteristics according to CD105 lo and CD105 hi .

Article Snippet: Unconjugated anti-CD105 mAb (clone K-ro23 and clone SN6, Santa Cruz Biotechnology, Dallas, TX) or isotype control (clone MPC-11, BD Biosciences, Heidelberg, Germany) as well as the comparative anti-CD105 clone 166707 (R&D Systems, Minneapolis, MN) and the respective isotype control (IgG1 isotype, clone MG1-45, BioLegend) were added at 5 nM, followed by a mouse-specific PE conjugate (1:100).

Techniques: