Journal: Analytical and Bioanalytical Chemistry

Article Title: MALDI imaging mass spectrometry of N -linked glycans on formalin-fixed paraffin-embedded murine kidney

doi: 10.1007/s00216-014-8293-7

Figure Lengend Snippet: MALDI imaging MS of N -linked glycans released from formalin-fixed murine kidney sections. Formalin-fixed murine kidney sections were treated with antigen retrieval prior to printing of 30 nL/spot dialyzed PNGase F or buffer control arrays with 250 μm spacing. 2,5-DHB (20 mg/mL) was sprayed onto the sections and MS spectra were acquired by oversampling at 100 μm intervals using a MALDI-TOF/TOF MS instrument. Data was loaded raw into SCiLS, preprocessed for baseline subtraction and normalization to total ion current (TIC) prior to segmentation analysis (maximum processing mode, interval width of 0.5 Da, strong smoothing) and pLSA (10 component, interval width of 0.5 Da, random initiation). Regions are outlined for PNGase F regions 1 ( red ) and 2 ( blue ), as well as control ( green ) and calibrant ( small red box ) regions. a The segmentation map for the entire data set, which discriminates between the cortex and medulla/pelvis of the kidney at the segmentation levels selected (see inset ). b – d Components 1, 2, and 8 from the pLSA analysis, which also discriminate the same regions of the tissue. e – h Ion intensity maps for (Hex) 2 (HexNAc) 2 + (Man) 3 (GlcNAc) 2 ( m / z 1663.632), (Hex) 6 + (Man) 3 (GlcNAc) 2 ( m / z 1905.697), (Hex)2(HexNAc) 3 (Deoxyhexose) 3 + (Man) 3 (GlcNAc) 2 ( m / z 2304.909), and (Hex) 3 (HexNAc) 4 (Deoxyhexose) 4 + (Man) 3 (GlcNAc) 2 ( m / z 2816.115). Intensity scales are autocorrected for these intensity maps (with weak denoising) and scale bars are included

Article Snippet: Abundance weighted mean (AWM, weighted by S / N ) m / z was calculated for each of these peak groups. (ii) MALDI imaging: Data was analyzed by SCiLS Lab (version 2014a, SCiLS GmbH, Bruker Daltonics) or by exporting peak lists from flexAnalysis (TopHat baseline subtraction and SN >3) and in-house R [ ] processing for difference in proportions statistic (DIPPS) analysis.

Techniques: Imaging, Control

Journal: Cancers

Article Title: DNA Replication Licensing Protein MCM10 Promotes Tumor Progression and Is a Novel Prognostic Biomarker and Potential Therapeutic Target in Breast Cancer

doi: 10.3390/cancers10090282

Figure Lengend Snippet: Cancer hallmarks of stable MCM10 knockdown (KD) in MCF 7 cells. ( A , B ) relative expression of MCM10 in stable MCM10-KD MCF 7 cell lines assessed by qPCR and Western blots ( n = 3, *** p < 0.001); ( C ) apoptosis analysis by Annexin V/Propidium Iodide flow cytometry showed no difference between stable MCM10-control MCF7 cell lines and stable MCM10-KD MCF 7 cell lines ( n = 3, * p < 0.05); ( D ) following MCM10 KD, the MCM10-KD MCF7 cells showed a decrease in cell proliferation monitored for seven days using phase contrast images in Incucyte ZOOM analysis software ( n = 6); ( E ) MTT assay using stable cells showed a similar observation indicating decreased proliferation in MCM10-KD MCF 7 cells ( n = 3, * p < 0.05); ( F ) analysis cell cycle markers by western blot and QPCR showed a decrease in cycling D1 expression indicating impaired cell proliferation in in MCM10-KD MCF 7 cells ( n = 3, * p < 0.05); ( G ) wound healing migration assay performed by using Incucyte ZOOM showed a decrease in cell migration in MCM10-KD MCF 7 cell lines. Wound heal quantified by relative Gap size using Incucyte ZOOM software ( n = 6, * p < 0.05); ( H ) Transwell cell migration assay was used to confirm our observation and showed a similar decrease in the number of migrating cells in MCM10-KD MCF 7 cells ( n = 3, ** p < 0.01); ( I ) colony formation assay performed using soft agar showed a decrease in the number of colonies in MCM10-KD MCF 7 cells compared to the control cells. ( n = 3, * p < 0.05).

Article Snippet: Data were analyzed using the IncuCyte Zoom software (version 2014a Essen Bioscience, Ann Arbor, MI, USA), which quantified cell surface area coverage as confluence values.

Techniques: Expressing, Western Blot, Flow Cytometry, Software, MTT Assay, Migration, Cell Migration Assay, Colony Assay