Journal: Neuron

Article Title: Location and Plasticity of the Sodium Spike Initiation Zone in Nociceptive Terminals In Vivo.

doi: 10.1016/j.neuron.2019.03.005

Figure Lengend Snippet: Figure 1. In Vivo Optical Recording from Nociceptive Corneal Nerve Terminal Endings and Distal Axons Showed Initiation and Propagation of Capsaicin-Induced Signaling and Their Dependence on VGCCs (A and B) Illustration of the experimental setup. An anesthetized mouse with the head stabilized and eye placed under an epifluorescence microscope (A). The puff of 0.5 mM capsaicin was applied above a superficial, ramifying terminal ending. The well-defined morphology of corneal innervation (B) allowed tracing of the terminal to its intraepithelial fibers, collating thereafter into subbasal and stromal plexuses. (C) Left: epifluorescence image of the recorded terminal taken at the corneal superficial epithelium surface. Orange arrow indicates the region of interest (ROI) from which the recordings were performed. The location of the puff pipette is outlined by dashed lines. Right and middle: representative traces of optical re- cordings from the terminal in the superficial epithelium following application of capsaicin before (middle, black trace) and after 60-min incubation with 50 mM benidipine (right, orange trace). (D) Boxplots and individual paired values of the terminal fluorescence intensities following application of capsaicin on the same terminal before and during in- cubation with benidipine. Comparison between ‘‘vehicle’’ and ‘‘benidipine’’ groups, p < 0.0001; one-sample t test; n = 14 terminals from 14 different eyes from 7 mice. (E) Same as in (C), but the image was taken and recordings were performed from an intraepithelial nerve fiber traced from a stimulated terminal. (F) Same as in (D) but for intraepithelial fibers. Comparison between vehicle and benidipine groups, p = 0.0005; one-sample t test; n = 6 fibers from 6 different eyes from 3 mice.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-neuronal class III b-tubulin Covance Research Products Cat#PRB-435P; RRID: AB_291637 Mouse monoclonal anti-Nav1.8 Abcam Cat#ab93616; RRID: AB_10561455 Goat anti-TRPV1 Neuromics Cat#GT15129; RRID: AB_2209002 Alexa Fluor 488 goat anti-rabbit IgG Molecular Probes ThermoFisher Cat#A11008; RRID: AB_143165 Alexa Fluor 555 goat anti-mouse IgG Molecular Probes ThermoFisher Cat#A32727; RRID: AB_2633276 Alexa Fluor 647 donkey anti-goat IgG Molecular Probes ThermoFisher Cat#A21447; RRID: AB_2535864 Bacterial and Virus Strains pENN.AAV.hSyn.TurboRFP.WPRE.RBG James M. Wilson Addgen AAV1; 105552-AAV1; RRID: Addgene_105552 pAAV.Syn.GCaMP6s.WPRE.SV40 Chen et al., 2013 Addgen AAV1; 100843-AAV1; RRID: Addgene_100843 Chemicals, Peptides, and Recombinant Proteins Oxybuprocaine hydrochloride 0.4% Fisher pharmaceutical Brand name: LOCALIN Benidipine hydrochloride Alomone Labs Cat#B-120 Anisomycin Sigma-Aldrich Cat#A9789 Recombinant Rat TNF-a Peprotech Cat#400-14 Recombinant Rat IL-1b Peprotech Cat#400-01B XE 991 dihydrochloride Tocris Cat#2000 Capsaicin Sigma-Aldrich Cat#M2028 Hyaluronidase type IV-S Sigma-Aldrich Cat#H3884 Hoechst 33342 Molecular Probes ThermoFisher Cat#H3570 Fluoromount-G Invitrogen ThermoFisher Cat#00-4958-02 Experimental Models: Organisms/Strains Mouse: C57BL/6 Envigo-Israel N/A Software and Algorithms Neuroplex RedShirt Imaging N/A Turbo-SM RedShirt Imaging N/A pCLAMP Molecular Devices Version 10.2 OriginPro OriginLab Corporation Version 6.0.0 CorelDraw Corel Corporation Version X6 Office Suite Microsoft Version 2016 MATLAB Mathworks Version R2016a NEURON terminal computational model Du et al., 2014; Barkai et al., 2017; This paper N/A Fiji ImageJ, NIH Version 1.4

Techniques: In Vivo, Microscopy, Transferring, Incubation, Comparison

Journal: International Journal of Molecular Medicine

Article Title: Role of ARID1A in epithelial-mesenchymal transition in breast cancer and its effect on cell sensitivity to 5FU

doi: 10.3892/ijmm.2020.4727

Figure Lengend Snippet: Association between ARID1A expression and the clinicopathological characteristics of patients with breast cancer.

Article Snippet: The pCMV6-XL4 and pCMV6-XL4-ARID1A (cat. no. SC303719) vectors were obtained from OriGene Technologies, Inc., and pCMV6-XL4 was used as a negative control (NC).

Techniques: Expressing

Journal: International Journal of Molecular Medicine

Article Title: Role of ARID1A in epithelial-mesenchymal transition in breast cancer and its effect on cell sensitivity to 5FU

doi: 10.3892/ijmm.2020.4727

Figure Lengend Snippet: Sequences of primers used for RT-qPCR.

Article Snippet: The pCMV6-XL4 and pCMV6-XL4-ARID1A (cat. no. SC303719) vectors were obtained from OriGene Technologies, Inc., and pCMV6-XL4 was used as a negative control (NC).

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: Role of ARID1A in epithelial-mesenchymal transition in breast cancer and its effect on cell sensitivity to 5FU

doi: 10.3892/ijmm.2020.4727

Figure Lengend Snippet: Association of ARID1A expression and breast cancer. (A) Detection of ARID1A expression in breast cancer tissues (n=3) and their matched adjacent tissues (n=3) by immunohistochemistry. The tissue sections are numbered from 1-3. (B) Measurement of the mRNA expression of ARID1A in tissues by RT-qPCR. (C) Detection of the mRNA expression of ARID1A in MCF-10A, BT20, BT474, T-47D, MCF7, MDA-MB-231 and MDA-MB-453 cells by RT-qPCR. (D) Statistical analysis of the relationship between the overall 5-year survival rate and the ARID1A expression level; the overall survival rate was calculated by Kaplan-Meier survival analysis and compared using the log-rank test. Data were obtained from 3 representative experiments and are presented as the means ± standard deviation. The experiment was repeated 3 times (n=3). ** P<0.001 vs. adjacent tissue or MCF-10A cells. ARID1A, AT-rich interactive domain 1A.

Article Snippet: The pCMV6-XL4 and pCMV6-XL4-ARID1A (cat. no. SC303719) vectors were obtained from OriGene Technologies, Inc., and pCMV6-XL4 was used as a negative control (NC).

Techniques: Expressing, Immunohistochemistry, Quantitative RT-PCR, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: Role of ARID1A in epithelial-mesenchymal transition in breast cancer and its effect on cell sensitivity to 5FU

doi: 10.3892/ijmm.2020.4727

Figure Lengend Snippet: Effects of function of ARID1A on the viability of breast cancer cells treated with 5-FU. (A and B) MCF7 cells were transfected with siARID1A and cultured for 48 h, and the transfection efficiency of siARID1A was detected by WB analysis. (C) The mRNA expression of ARID1A in MCF7 cells was detected by RT-qPCR after the cells were transfected with siARID1A. (D and E) MDA-MB-231 cells were transfected with ARID1A overexpression vector and cultured for 48 h, and the transfection efficiency of ARID1A was detected by WB analysis. (F) The mRNA expression of ARID1A in MDA-MB-231 cells was detected by RT-qPCR after the cells were transfected with ARID1A overexpression vector. (G) The effect of ARID1A silencing on the viability of MCF-7 cells treated with various concentrations of 5-FU (5, 10, 20, 40 and 80 µ g/ml) for 24 h was detected by proliferation assay. (H) Effect of ARID1A overexpres-sion on the viability of MDA-MB-231 cells treated with various concentrations of 5-FU (5, 10, 20, 40 and 80 µ g/ml) for 24 h was detected by proliferation assay. (I) The mRNA expression of ARID1A in MCF-7 cells treated with various concentrations of 5-FU (5, 10, 20, 40 and 80 µ g/ml) and siARID1A was detected by RT-qPCR. (J) The mRNA expression of ARID1A in MDA-MB-231 cells treated with various concentrations of 5-FU (5, 10, 20, 40 and 80 µ g/ml) and pCMV6-XL4-ARID1A was detected by RT-qPCR. Data were obtained from 3 representative experiments and are presented as the means ± standard deviation the experiment was repeated 3 times (n=3). * P<0.05, ** P<0.001 vs. siNC or NC; ## P<0.001 vs. 5 + siNC or 5 + NC; ^ P<0.05, ^^ P<0.001 vs. 10 + siNC or 10 + NC; Δ P<0.05, ΔΔ P<0.001 vs. 20 + siNC or 20 + NC; ΦΦ P<0.001 vs. 40 + siNC or 40 + NC; Ψ P<0.05, ΨΨ P<0.001 vs. 80 + siNC or 80 + NC. ARID1A, AT-rich interactive domain 1A; 5-FU, 5-fluorouracil; WB analysis, western blot analysis.

Article Snippet: The pCMV6-XL4 and pCMV6-XL4-ARID1A (cat. no. SC303719) vectors were obtained from OriGene Technologies, Inc., and pCMV6-XL4 was used as a negative control (NC).

Techniques: Transfection, Cell Culture, Expressing, Quantitative RT-PCR, Over Expression, Plasmid Preparation, Proliferation Assay, Standard Deviation, Western Blot

Journal: International Journal of Molecular Medicine

Article Title: Role of ARID1A in epithelial-mesenchymal transition in breast cancer and its effect on cell sensitivity to 5FU

doi: 10.3892/ijmm.2020.4727

Figure Lengend Snippet: Effects of ARID1A on the apoptosis and cell cycle of breast cancer cells treated with 5-FU. (A and B) The apoptosis of MCF7 and MDA-MB-231 cells following transfection with siARID1A or pCMV6-XL4-ARID1A and treatment with 40 µ g/ml 5-FU for 48 h was detected by flow cytometry. (C and D) The cell cycle of MCF7 and MDA-MB-231 cells following transfection with siARID1A or pCMV6-XL4-ARID1A and treatment with 40 µ g/ml 5-FU was detected by flow cytometry. Data were obtained from 3 representative experiments and are presented as the means ± standard deviation. The experiment was repeated 3 times (n=3). ** P<0.001 vs. siNC or NC; ## P<0.001 vs. siARID1A or ARID1A; ^ P<0.05, ^^ P<0.001 vs. siNC + 5-FU or NC + 5-FU. ARID1A, AT-rich interactive domain 1A; 5-FU, 5-fluorouracil.

Article Snippet: The pCMV6-XL4 and pCMV6-XL4-ARID1A (cat. no. SC303719) vectors were obtained from OriGene Technologies, Inc., and pCMV6-XL4 was used as a negative control (NC).

Techniques: Transfection, Flow Cytometry, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: Role of ARID1A in epithelial-mesenchymal transition in breast cancer and its effect on cell sensitivity to 5FU

doi: 10.3892/ijmm.2020.4727

Figure Lengend Snippet: Effets of ARID1A on the migration, invasion and angiogenesis of breast cancer cells treated with 5-FU. (A and B) Detection of the migration rates of MCF7 and MDA-MB-231 cells following transfection with siARID1A or pCMV6-XL4-ARID1A and treatment with 40 µ g/ml 5-FU by wound healing migration assay at 0 and 24 h. (C and D) Measurement of the invasion rates of MCF7 and MDA-MB-231 cells following transfection with siARID1A or pCMV6-XL4-ARID1A and treatment with 40 µ g/ml 5-FU by invasion assay at 24 h. (E and F) Detection of the angiogenesis formation rates of MCF7 and MDA-MB-231 cells following transfection with siARID1A or pCMV6-XL4-ARID1A and treatment with 40 µ g/ml 5-FU by tube formation assay. Data were obtained from 3 representative experiments and are presented as the means ± standard deviation. The experiment was repeated 3 times (n=3). ** P<0.001 vs. siNC or NC; ## P<0.001 vs. siARID1A or ARID1A; ^ P<0.05, ^^ P<0.001 vs. siNC + 5-FU or NC + 5-FU. ARID1A, AT-rich interactive domain 1A; 5-FU, 5-fluorouracil.

Article Snippet: The pCMV6-XL4 and pCMV6-XL4-ARID1A (cat. no. SC303719) vectors were obtained from OriGene Technologies, Inc., and pCMV6-XL4 was used as a negative control (NC).

Techniques: Migration, Transfection, Invasion Assay, Tube Formation Assay, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: Role of ARID1A in epithelial-mesenchymal transition in breast cancer and its effect on cell sensitivity to 5FU

doi: 10.3892/ijmm.2020.4727

Figure Lengend Snippet: Effects of ARID1A on the expression of EMT-related proteins and mRNAs. (A and B) Detection of the protein expression of E-cadherin, N-cadherin and Vimentin in MCF7 cells by WB analysis. (C) Detection of the mRNA expression of E-cadherin, N-cadherin and Vimentin in MCF7 cells by RT-qPCR. (D and E) Detection of the protein expression of E-cadherin, N-cadherin and Vimentin in MDA-MB-231 cells by WB analysis. (F) The detection of the mRNA expressions of E-cadherin, N-cadherin and Vimentin in MDA-MB-231 cells by RT-qPCR. Data were obtained from 3 representative experiments and are presented as the means ± standard deviation. The experiment was repeated 3 times (n=3). ** P<0.001 vs. siNC or NC; ## P<0.001 vs. siARID1A or ARID1A; ^^ P<0.001 vs. siNC + 5-FU or NC + 5-FU. ARID1A, AT-rich interactive domain 1A; 5-FU, 5-fluorouracil; WB analysis, western blot analysis.

Article Snippet: The pCMV6-XL4 and pCMV6-XL4-ARID1A (cat. no. SC303719) vectors were obtained from OriGene Technologies, Inc., and pCMV6-XL4 was used as a negative control (NC).

Techniques: Expressing, Quantitative RT-PCR, Standard Deviation, Western Blot

Journal: International Journal of Molecular Medicine

Article Title: Role of ARID1A in epithelial-mesenchymal transition in breast cancer and its effect on cell sensitivity to 5FU

doi: 10.3892/ijmm.2020.4727

Figure Lengend Snippet: Effects of ARID1A on the cell cycle, apoptosis, and the expression of angiogenesis-related proteins and mRNAs. (A and B) Detection of the protein expression of ARID1A, VEGF, cyclin D1, Bcl-2 and Bax in MCF7 cells by WB analysis. (C) Detection of the mRNA expressions of ARID1A, VEGF, cyclin D1, Bcl-2 and Bax in MCF7 cells by RT-qPCR. (D and E) Detection of the protein expression of ARID1A, VEGF, cyclin D1, Bcl-2 and Bax in MDA-MB-231 cells by WB analysis. (F) Detection of the mRNA expression of ARID1A, VEGF, cyclin D1, Bcl-2 and Bax in MDA-MB-231 cells by RT-qPCR. Data were obtained from 3 representative experiments and are presented as the means ± standard deviation. The experiment was repeated 3 times (n=3). ** P<0.001 vs. siNC or NC; ## P<0.001 vs. siARID1A or ARID1A; ^ P<0.05, ^^ P<0.001 vs. siNC + 5-FU or NC + 5-FU. ARID1A, AT-rich interactive domain 1A; 5-FU, 5-fluorouracil; WB analysis, western blot analysis.

Article Snippet: The pCMV6-XL4 and pCMV6-XL4-ARID1A (cat. no. SC303719) vectors were obtained from OriGene Technologies, Inc., and pCMV6-XL4 was used as a negative control (NC).

Techniques: Expressing, Quantitative RT-PCR, Standard Deviation, Western Blot