deparaffinization rehydration  (Agilent technologies)

Journal: Cancer discovery

Article Title: Evolution of neoantigen landscape during immune checkpoint blockade in non-small cell lung cancer

doi: 10.1158/2159-8290.CD-16-0828

Figure Lengend Snippet: Emergence of resistance to immune checkpoint blockade is associated with elimination of mutation associated neoantigens by loss of heterozygosity and a more diverse T-cell repertoire independent of PD-L1 expression Panel A shows computed tomographic (CT) images of patient CGLU117 at baseline, at the time of therapeutic response and at time of acquired resistance. Pre-treatment CT image of the abdomen, demonstrates a right adrenal mass (T1, circled), radiologic tumor regression is noted after 2 months of treatment, followed by disease relapse at 4 months from treatment initiation with a markedly increased right adrenal metastasis (T2, circled). 3 rd follow up CT demonstrates further disease progression in the adrenal lesion. Tumor burden kinetics for target lesions by RECIST criteria are shown in panel B. Peripheral T cell expansion of a subset of intratumoral clones was noted to peak at the time of response and decrease to baseline levels at the time of resistance (panel C). Productive TCR frequency denotes the frequency of a specific rearrangement that can produce a functional protein receptor among all productive rearrangements. Panels D and E show B allele frequency graphs for chromosome 17, a value of 0.5 indicates a heterozygous genotype whereas allelic imbalance is observed as a deviation from 0.5. The region that undergoes LOH in the resistant tumor (panel E, orange box) contains 3 mutation associated neoantigens that are thus eliminated. No differences in CD8+ T cell density (panel F, G) or PD-L1 expression (panel H, I) were observed between baseline and resistant tumors.

Article Snippet: Similarly, slides were deparaffinized, rehydrated, antigen retrieved and incubated with a mouse anti-human CD8 antibody (Dako, CA) diluted 1:100 overnight at 4°C, followed by a 30-minute incubation with the FLEX+ polymer system.

Techniques: Mutagenesis, Expressing, Clone Assay, Functional Assay

Journal: Endocrinology

Article Title: Diminished Pancreatic ?-Cell Mass in Securin-Null Mice Is Caused by ?-Cell Apoptosis and Senescence

doi: 10.1210/en.2008-0972

Figure Lengend Snippet: p21 up-regulation in PTTG −/− mice. Paraffin sections of pancreas from WT or PTTG −/− mouse embryo at embryonic d 17 (ED17), newborn (NB), and 1- to 8-wk-old mice were stained with guinea pig antiinsulin (Ins; red ) or rabbit

Article Snippet: For staining of islet hormones, 5-μm paraffin pancreatic sections were deparaffinized, rehydrated, and incubated with guinea pig antiinsulin at 1:200 to 1:2000 dilution, rabbit antiglucagon, antisomatostatin, or antipancreatic polypeptide (Dako, Carpinteria, CA) at 1:200 dilution, followed by rhodamine-labeled antiguinea pig or fluorescein isothiocyanate (FITC)-labeled antirabbit antiserum.

Techniques: Mouse Assay, Staining

Journal: Scientific Reports

Article Title: The Role of Inflammation in β-cell Dedifferentiation

doi: 10.1038/s41598-017-06731-w

Figure Lengend Snippet: Pancreatic islets of patients with type 2 diabetes contain more CD45 + immune cells than islets of non-diabetic subjects. ( a ) Representative islet histology of type 2 diabetic (T2D) and non-diabetic (ND) subjects. Peri-islet and intra-islet CD45 + immune cell count ( b and c , respectively) and distribution ( d and e , respectively) in human pancreatic tissue sections of T2D and ND control subjects. Islet area ( f ) and islet-area-corrected average CD45 + immune cell count ( g ) in T2D and ND control subjects. ( b – e ) n = 17, 16 for T2D and ND subjects, respectively, ( f , g ) n = 14, 10 for T2D and ND subjects. n = 30–35 islets/subject were counted. ***P

Article Snippet: Tissue slides were deparaffinized, rehydrated and stained with mouse anti-human CD45 (1:100, overnight at 4 °C; DAKO, M0701) and biotinylated anti-mouse immunoglobulins (1:200, 60 min at room temperature, DAKO, E0354) and visualized with DAB (2 min, DAKO, K3467).

Techniques: Cell Counting

Journal: PLoS ONE

Article Title: Radiotherapy resistance acquisition in Glioblastoma. Role of SOCS1 and SOCS3

doi: 10.1371/journal.pone.0212581

Figure Lengend Snippet: Molecular features of stablished GBM cell lines. Electropherograms of IDH1 (a , f , k , p, u, aa ), IDH2 ( b , g , l , q , w , ab ), and TERT ( c , h , m , r , x , ac ), and immunocytochemistry images of P53 ( d , i , n , s , y , ad ) and PTEN ( e , j , o , t , z , ae ) of all cell lines are shown. The first line corresponds to GB16, the second line to GB37, the third line to GB39, the fourth line to GB40, the fifth line to GB42 and the sixth line to GB48. Red rectangles show the codon which harbours the mutation. The images were made to 40ꓫ.

Article Snippet: Then, sections were deparaffined, rehydrated and incubated with primary antibodies anti-P53 (DO-7, Dako) or anti-PTEN (6H2.1, Dako).

Techniques: Immunocytochemistry, Mutagenesis