Journal: Journal of Cancer

Article Title: MCMs expression in lung cancer: implication of prognostic significance

doi: 10.7150/jca.20777

Figure Lengend Snippet: Expression status of MCM2, MCM5 and MCM6 in NSCLC tissue samples Representative immunohistochemical microphotographs of MCM2 (a,b,c), MCM5 (d,e,f) and MCM6 (g,h,i) with high (positive) and low (negative) expression in NSCLC and their adjacent non-malignant tissues. The subtypes are SCCs (b, e, h) and ADCs (c, f, i). Bar = 100μm.

Article Snippet: The slides were blocked with 10% normal goat serum for 30 min at 37°C and washed and incubated overnight at 4°C with mouse monoclonal antibody against MCM2 (1:200; 10513-1-AP, Proteintech Group, Inc. Chicago, IL, USA), MCM5 (1:200; 11703-1-AP, Proteintech Group, Inc. Chicago, IL, USA), or MCM6 (1:200; 13347-2-AP, Proteintech Group, Inc. Chicago, IL, USA).

Techniques: Expressing, Immunohistochemical staining

Journal: Journal of Cancer

Article Title: MCMs expression in lung cancer: implication of prognostic significance

doi: 10.7150/jca.20777

Figure Lengend Snippet: Relationship Between Protein Overexpression and Clinicopathologic Parameters

Article Snippet: The slides were blocked with 10% normal goat serum for 30 min at 37°C and washed and incubated overnight at 4°C with mouse monoclonal antibody against MCM2 (1:200; 10513-1-AP, Proteintech Group, Inc. Chicago, IL, USA), MCM5 (1:200; 11703-1-AP, Proteintech Group, Inc. Chicago, IL, USA), or MCM6 (1:200; 13347-2-AP, Proteintech Group, Inc. Chicago, IL, USA).

Techniques: Over Expression

Journal: Journal of Cancer

Article Title: MCMs expression in lung cancer: implication of prognostic significance

doi: 10.7150/jca.20777

Figure Lengend Snippet: Correlation of MCM2, MCM5 and MCM6 LIs with NSCLC patients' overall survival (OS) Kaplan-Meier curves showing the association between LIs of MCM2 (A), MCM5 (B), MCM6 (C) and OS in different stages and in different histological tumor types in tissue samples. All the P values are shown in the graph, by log-rank test.

Article Snippet: The slides were blocked with 10% normal goat serum for 30 min at 37°C and washed and incubated overnight at 4°C with mouse monoclonal antibody against MCM2 (1:200; 10513-1-AP, Proteintech Group, Inc. Chicago, IL, USA), MCM5 (1:200; 11703-1-AP, Proteintech Group, Inc. Chicago, IL, USA), or MCM6 (1:200; 13347-2-AP, Proteintech Group, Inc. Chicago, IL, USA).

Techniques:

Journal: Journal of Cancer

Article Title: MCMs expression in lung cancer: implication of prognostic significance

doi: 10.7150/jca.20777

Figure Lengend Snippet: Univariate and multivariate analysis of survival in patients with squamous cell carcinoma

Article Snippet: The slides were blocked with 10% normal goat serum for 30 min at 37°C and washed and incubated overnight at 4°C with mouse monoclonal antibody against MCM2 (1:200; 10513-1-AP, Proteintech Group, Inc. Chicago, IL, USA), MCM5 (1:200; 11703-1-AP, Proteintech Group, Inc. Chicago, IL, USA), or MCM6 (1:200; 13347-2-AP, Proteintech Group, Inc. Chicago, IL, USA).

Techniques:

Journal: Journal of Cancer

Article Title: MCMs expression in lung cancer: implication of prognostic significance

doi: 10.7150/jca.20777

Figure Lengend Snippet: Overexpression of MCM2, MCM5 and MCM6 mRNAs in NSCLC samples Transcript levels of MCM2, MCM5 and MCM6 in non-tumor tissues (N) and NSCLC tissues (SCCs or ADCs) from TCGA_LUSC (N=553) and TCGA_LUAD (N=571) datasets. Data are represented as mean±SD. All P values were calculated by t-test analysis, *** P <0.001.

Article Snippet: The slides were blocked with 10% normal goat serum for 30 min at 37°C and washed and incubated overnight at 4°C with mouse monoclonal antibody against MCM2 (1:200; 10513-1-AP, Proteintech Group, Inc. Chicago, IL, USA), MCM5 (1:200; 11703-1-AP, Proteintech Group, Inc. Chicago, IL, USA), or MCM6 (1:200; 13347-2-AP, Proteintech Group, Inc. Chicago, IL, USA).

Techniques: Over Expression

Journal: Journal of Cancer

Article Title: MCMs expression in lung cancer: implication of prognostic significance

doi: 10.7150/jca.20777

Figure Lengend Snippet: Correlation of MCM2, MCM5 and MCM6 mRNAs with patients' overall survival (OS) in TCGA database. The survival analysis stratified by MCM2, MCM5 and MCM6 expression levels in datasets LUAD - TCGA - Lung adenocarcinoma June 2016 (N=475) and LUSC - TCGA - Lung squamous cell carcinoma June 2016 (N=175) were analyzed using the Survexpress online platforms. The Log-Rank P values are shown in the graph.

Article Snippet: The slides were blocked with 10% normal goat serum for 30 min at 37°C and washed and incubated overnight at 4°C with mouse monoclonal antibody against MCM2 (1:200; 10513-1-AP, Proteintech Group, Inc. Chicago, IL, USA), MCM5 (1:200; 11703-1-AP, Proteintech Group, Inc. Chicago, IL, USA), or MCM6 (1:200; 13347-2-AP, Proteintech Group, Inc. Chicago, IL, USA).

Techniques: Expressing

Journal: PLoS Genetics

Article Title: Selective role of the DNA helicase Mcm5 in BMP retrograde signaling during Drosophila neuronal differentiation

doi: 10.1371/journal.pgen.1010255

Figure Lengend Snippet: (A-C) Schematic representation of Drosophila embryo Nervous System (A), with detailed position of the Ap cluster within the VNC (B) and some of the most representative expressed markers which identifies each Tv neuron within the Ap Cluster (C). (D-F) Immunostaining for proFMRFa in Control (D), Mcm5 mutant (E), Mcm5 misexpression (F), at stage 18 AEL (Scale bar 20μm). (E) Quantification of proFMRFa expressing cells within the Ap cluster in Control, Mcm5 mutant and Mcm5 misexpression. (G-H) Quantification of proFMRFa expressing cells within the thoracic and second suboesophageal segment (SE2) in control, Mcm5 mutant and Mcm5 misexpression (U-Mann-Whitney test; n≥10 CNS per genotype; *** = p<0,001). (I-J) Immunostaining for Nplp1 in Control (G) and Mcm5 mutant (H), at stage 18h AEL. Genotypes: (A, G, I) Oregon-R , w 1118 . (B, H, J) Mcm5 exc222 /Mcm5 exc222 (C) prospero-Gal4/UAS-Mcm5 .

Article Snippet: Mcm5 antibody were raised in Guinea Pigs against the nh2- C+SSNKSAPSEPAEGEI–conh2w and nh2- C+FGRWDDTKGEENIDF–conh2 peptides (Cultek, Madrid, Spain).

Techniques: Immunostaining, Mutagenesis, Expressing, MANN-WHITNEY

Journal: PLoS Genetics

Article Title: Selective role of the DNA helicase Mcm5 in BMP retrograde signaling during Drosophila neuronal differentiation

doi: 10.1371/journal.pgen.1010255

Figure Lengend Snippet: Gene screen of FMRFa altered expression pattern. Values relative to the gene (FlyBase gene symbol) identifying the loss of FMRFa in Tv4 cells scored by immunostainig of proFMRFa in embryos at stages 11–16. It is indicated the specific allele tested as well as the fly stock origin.

Article Snippet: Mcm5 antibody were raised in Guinea Pigs against the nh2- C+SSNKSAPSEPAEGEI–conh2w and nh2- C+FGRWDDTKGEENIDF–conh2 peptides (Cultek, Madrid, Spain).

Techniques: Expressing

Journal: PLoS Genetics

Article Title: Selective role of the DNA helicase Mcm5 in BMP retrograde signaling during Drosophila neuronal differentiation

doi: 10.1371/journal.pgen.1010255

Figure Lengend Snippet: (A-B) Expression of the temporal factors Castor (Cas) and Grainy head (Grh) in the Ap cluster in control and Mcm5 mutants, at St16. Eya was used to visualize Ap neurons. (E-J) Expression of Ap neuron determinants Nab, Squeeze (Sqz), Dimmed (Dimm) and phosphorylated form of Mothers against dpp (pMad) in control and Mcm5 mutants, at stage 18 AEL. Eya was used to visualize Ap neurons. (K-N) Expression of ap based on the ap-Gal4>UAS-GFP reporter expression, Dachshund (Dac) and Seven up (Svp) in control and Mcm5 mutants, at stage AFT. Merge and individual antibody images are shown in each panel. Representative images of n≥3 CNS per genotype, n≥ 18 hemisegments per genotype. Genotypes: (A, C, E, G, I, K, M) w 1118 . (B, D, F, H, J, L, N) Mcm5 exc222 /Mcm5 exc222 (Scale bar 5μm).

Article Snippet: Mcm5 antibody were raised in Guinea Pigs against the nh2- C+SSNKSAPSEPAEGEI–conh2w and nh2- C+FGRWDDTKGEENIDF–conh2 peptides (Cultek, Madrid, Spain).

Techniques: Expressing

Journal: PLoS Genetics

Article Title: Selective role of the DNA helicase Mcm5 in BMP retrograde signaling during Drosophila neuronal differentiation

doi: 10.1371/journal.pgen.1010255

Figure Lengend Snippet: (A-E) Expression of Eya and proFMRFa, in control (A), Mcm5 mutants (B), expression of Gbb ligand ( gbb ) in Mcm5 (C), expression of the activated type I BMP receptors ( sax A and tkv A ) in Mcm5 mutants (D), expression of wild type forms of the type I BMP receptors ( sax and tkv ) in Mcm5 mutants (E), at stage 18h AEL (thoracic segments T1-T3). (F) Quantification of proFMR expressing cells (Tv4) in control, Mcm5 mutants and rescue genotypes (U-Mann-Whitney test; n≥6 CNS per genotype, n≥ 30 hemisegments per genotype, *** = p < 0.001; n.s = non-significant). Merge and individual antibody images are shown in each panel. Genotypes: (A) Oregon-R , w 1118 . (B) pros-Gal4 ; Mcm5 exc222 . (C) pros-Gal4/UAS-gbb ; Mcm5 exc222 (D) pros-Gal4/UAS-saxA , UAS-tkvA ; Mcm5 exc222 . (E) pros-Gal4/UAS-sax , UAS-tkv ; Mcm5 exc222 .

Article Snippet: Mcm5 antibody were raised in Guinea Pigs against the nh2- C+SSNKSAPSEPAEGEI–conh2w and nh2- C+FGRWDDTKGEENIDF–conh2 peptides (Cultek, Madrid, Spain).

Techniques: Expressing, MANN-WHITNEY

Journal: PLoS Genetics

Article Title: Selective role of the DNA helicase Mcm5 in BMP retrograde signaling during Drosophila neuronal differentiation

doi: 10.1371/journal.pgen.1010255

Figure Lengend Snippet: (A) Bland-Altman graph showing differences in the expression of every gene analyzed through RNA-seq from control and Mcm5 mutants in St16-AFT embryos. The expression of every gene is represented by a dot. Dots are located in the graph according to change in expression between conditions in a logarithmic scale, when compared with the media of the normalized expression. Genes with differences in expression between genotypes are represented in red, otherwise they are grey. Genes represented in positive values show higher expression in Mcm5 mutants than in controls and vice versa (graph made with DESeq2, statistical analysis performed by hypergeometric distribution adjusted by Benjamini-Hochberg; p-value < 0,05). (B) Fold change histogram of the relative expression in Mcm5 mutants compared to the control of representative genes involved in Tv4 specification, ap , eya , nab , sqz , dimm , dac , svp , and genes involved in BMP signaling pathway, gbb , Mad , sax and tkv . (C) Alignment distribution of reads obtained after RNA-seq analysis of tkv gene in control and Mcm5 mutants. The top of the graph shows the enrichment in sequences corresponding to every exonic region and the gene sequence of tkv gene represented in indigo (TopHat2) and blue (Flybase). A striking reduction in read coverage for the exons 1 and 2 (black arrows) of the tkv gene in Mcm5 mutants comparing to the control is shown. On the bottom, the graph shows the different isoforms of tkv RNAm represented in magenta, other genes located in the same chromosome region in green (its RNAm in yellow), and non-coding RNAs in orange (Flybase) (graph made with TopHat2, IGV). Genotypes: (A, B) w 1118 and Mcm5 exc222 /Mcm5 exc222 .

Article Snippet: Mcm5 antibody were raised in Guinea Pigs against the nh2- C+SSNKSAPSEPAEGEI–conh2w and nh2- C+FGRWDDTKGEENIDF–conh2 peptides (Cultek, Madrid, Spain).

Techniques: Expressing, RNA Sequencing Assay, Sequencing

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Similarities and differences between rat and mouse chondrocyte gene expression induced by IL-1β

doi: 10.1186/s13018-021-02889-2

Figure Lengend Snippet: Proliferation-related DEGs in the RM group and MM group. A The expression level of MCM2 was detected by immunocytochemistry in rat and mouse chondrocytes after IL-1β intervention. B In situ expression of MCM5 was evaluated by cell immunofluorescence (green). The nuclei were counterstained by DAPI (blue). C Lamin B1, PCNA and Cyclin D1 expression levels were assessed by Western blots. GAPDH was used as the internal control. * P < 0.05; ** P < 0.001

Article Snippet: Thereafter, the cells were incubated in PBST with 10% BSA for 60 min to block nonspecific antibody binding, incubated with primary MCM5 antibody (A5489, Bimake, China) in 1% BSA for 60 min at RT, washed with PBS three times, and incubated with a secondary antibody in 1% BSA for 30 min at RT.

Techniques: Expressing, Immunocytochemistry, In Situ, Immunofluorescence, Western Blot, Control