Journal: Scientific Reports

Article Title: Water adsorption on TiO 2 surfaces probed by soft X-ray spectroscopies: bulk materials vs. isolated nanoparticles

doi: 10.1038/srep15088

Figure Lengend Snippet: ( a ) Valence spectra obtained with a suspension of solvated NPs sprayed out with an atomizer ( blue ) and on “as-received” dry NPs sprayed out with an aerosoliser ( green ) at 100 eV photon energy. ( b ) Comparison of the edge of the valence band (4–12 eV) for “as-received” dry NPs ( green ) and hydrated NPs ( blue ). The spectra have been normalized relative to the top of the valence band.

Article Snippet: The second method was based on the hydration of a dry particles beam generated with a commercial aerosoliser ( Naneum Aerosolizer PA100S ).

Techniques: Suspension, Comparison

Journal: Scientific Reports

Article Title: Defective membrane repair machinery impairs survival of invasive cancer cells

doi: 10.1038/s41598-020-77902-5

Figure Lengend Snippet: Expression and subcellular localization of endogenous Anx in MDA-MB-231. ( a ) 10 µg of protein extracts from MDA-MB-231 and BeWo cells were analyzed by western-blot. The gel analysis plugin of ImageJ was used to quantify the density of bands. The histogram presents mean values (± SD) of the ratio Anx/GAPDH from ten independent experiments. Student t Test for independent samples. ** p < 0.01. ( b ) Revelation of AnxA5 in MDA-MB-231 and BeWo cells compared to 50 ng purified recombinant AnxA5 (Pure A5), presented as an example. The whole membrane, together with representative western-blot analysis for AnxA1, A2, A4 and A6, are presented in Supplementary Fig. . ( c ) MDA-MB-231 cells were immunostained for AnxA1, AnxA2, AnxA4, AnxA5 and AnxA6 (green), as indicated, and counterstained with DAPI (blue). The right-hand column presents magnified images extracted from the “Anx” column (indicated by inserts). Scale bar: 40 µm. ( d ) Quantification of fluorescence intensities after immunostaining of endogenous Anx in MDA-MB-231 cells. Fluorescence intensities were measured from 8-bit images by drawing a circular ROI inside the cytoplasm and measuring the mean pixel value. Mean pixel values from at least 20 cells are presented. Horizontal bars represent median values. The variance value is 323, 268, 224, 2372 and 2909 for AnxA1, A2, A4, A5 and A6, respectively. The coefficient of variation is 14%, 13%, 24%, 35% and 47% for AnxA1, A2, A4, A5 and A6, respectively. The expression level of AnxA5 or AnxA6 is heterogenous from one MDA-MB-231 cell to another. In some cells, AnxA5 and AnxA6 are expressed at a very low level.

Article Snippet: Semi-dry electrophoretic transfer (Bio-Rad, Hercules, CA, USA) onto PVDF membrane was performed for 1 h at 100 V. The cellular content in AnxA1 (37 kDa), AnxA2 (36 kDa), AnxA4 (32 kDa), AnxA5 (35 kDa), AnxA6 (68 kDa), and glyceraldehyde-3-phosphate deshydrogenase (GAPDH, loading control, 37 kDa) was detected with rabbit anti-AnxA1 polyclonal antibody (PA1006, BosterBio, Pleasanton, CA, USA), mouse anti-AnxA2 monoclonal antibody (3E8-B6, Sigma, Saint-Louis, MO, USA), mouse anti-AnxA4 monoclonal antibody (SAB4200121, Sigma), mouse anti-AnxA5 monoclonal antibody (AN5, Sigma), mouse anti-AnxA6 monoclonal antibody (sc-271859, Santa cruz Biotechnology), and rabbit anti-GAPDH polyclonal antibody (FL-335, Santa Cruz Biotechnology), respectively.

Techniques: Expressing, Western Blot, Purification, Recombinant, Membrane, Fluorescence, Immunostaining

Journal: International Journal of Cancer

Article Title: Cancer‐associated fibroblasts promote PD‐L1 expression in mice cancer cells via secreting CXCL5

doi: 10.1002/ijc.32278

Figure Lengend Snippet: CAFs‐derived CXCL5 promotes PD‐L1 expression in cancer cells. ( a ) The expression of PD‐L1 in B16, CT26, A375 and HCT116 was determined by flow cytometry after treated with different concentrations of recombined CXCL5. ( b ) The mRNA expression of PD‐L1 in B16, CT26, A375 and HCT116 was detected by RT‐PCR. * p < 0.05. ( c ) Western blot showed the protein levels of CXCR2 in B16 and CT26 after transfected with siCXCR2. ( d ) Flow cytometry showed the expression of PD‐L1 in B16 and CT26 cocultured with CAFs after transfected with siCXCR2. ( e ) The protein levels of CXCR2 in A375 and HCT116 after transfected with siCXCR2 were analyzed by western blotting. ( f ) The expression of PD‐L1 in A375 and HCT116 treated with CXCL5 after transfected with siCXCR2 was determined by flow cytometry. ( g ) CXCL5 and α‐SMA expression in 3T3 cells were detected by western after transfected with CXCL5‐overexpression plasmid. ( h ) PD‐L1 protein expression in B16 and CT26 cocultured with/without 3T3‐CXCL5 cells was analyzed by western blot and the normalized protein expression was analyzed by Image J. * p < 0.05.

Article Snippet: The following antibodies were used for IHC, flow cytometry and western blotting: rabbit monoclonal anti‐PD‐L1 (1:200, ab205921, Abcam, Cambridge, MA), mouse monoclonal anti‐α‐smooth muscle actin (α‐SMA; 1:200, BM0002, BOSTER Biological Technology, Pleasanton, CA), rabbit polyclonal anti‐CXCR2 (1:200, BA0732‐2, BOSTER).

Techniques: Derivative Assay, Expressing, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Over Expression, Plasmid Preparation

Journal: International Journal of Cancer

Article Title: Cancer‐associated fibroblasts promote PD‐L1 expression in mice cancer cells via secreting CXCL5

doi: 10.1002/ijc.32278

Figure Lengend Snippet: The correlation of CXCR2, PD‐L1 and p‐AKT in tumor tissues in vivo . ( a ) The expression level of CXCR2 and PD‐L1 in human melanoma and CRC tissues were detected by IHC and evaluated. ( b ) The correlation of CXCR2 and PD‐L1 in xenograft tumor tissues was analyzed by IHC and evaluated. ( c ) The correlation of p‐AKT and PD‐L1 in xenograft tumor tissues was detected by IHC and quantized. ( d ) Representative immunostaining images of PD‐L1, CXCR2 and p‐AKT in B16 and CT26 xenograft tumor tissues. Scale bar, 200 μm.

Article Snippet: The following antibodies were used for IHC, flow cytometry and western blotting: rabbit monoclonal anti‐PD‐L1 (1:200, ab205921, Abcam, Cambridge, MA), mouse monoclonal anti‐α‐smooth muscle actin (α‐SMA; 1:200, BM0002, BOSTER Biological Technology, Pleasanton, CA), rabbit polyclonal anti‐CXCR2 (1:200, BA0732‐2, BOSTER).

Techniques: In Vivo, Expressing, Immunostaining