Journal: Cell

Article Title: A novel class of ER membrane proteins regulates ER-associated endosome fission

doi: 10.1016/j.cell.2018.08.030

Figure Lengend Snippet: Key Resources Table

Article Snippet: Primary antibody concentrations were used as follows: TMCC1 (rabbit) ( Zhang et al., 2014 ) 1:1000; FAM21 (rabbit) 1:1000 (gift from Billadeau, Gomez and Billadeau 2009 ); VPS35 (rabbit) 1:3000 (Abcam ab97545), and CORO1C (rabbit) 1:2000 (Proteintech 14749-1-Ap); M6PR 2G11 (mouse) (ThermoFisher Ma 1-066), 1:1000; GAPDH (rabbit) 1:30,000 (Sigma-Aldrich G9545, St. Louis, MO).

Techniques: Injection, Purification, Affinity Purification, Recombinant, BIA-KA, Mutagenesis, Binding Assay, Sequencing, Software

Journal: Cellular & Molecular Biology Letters

Article Title: MiR-206 may suppress non-small lung cancer metastasis by targeting CORO1C

doi: 10.1186/s11658-020-00216-x

Figure Lengend Snippet: MiR-206 directly targets the 3′-UTR of CORO1C. a The sequence of human miR-206 and the predicted binding sites with miR-206 within the CORO1C 3′-UTR are shown. b MiR-206 suppresses CORO1C expression in A549 cells. The A549 cells were treated with NC mimic or miR-206 mimic for 24 h, and CORO1C expression was evaluated by western blotting assay. β-actin was used as a loading control. c MiR-206 represses CORO1C mRNA in A549 cells. The A549 cells were cotransfected with luciferase plasmids containing wild-type (WT) CORO1C 3′UTR or mutant-type (Mut) CORO1C 3′UTR. The cells were also treated with miR-206 mimic at the same time. The cells were lysed to measure the relative luciferase activity. Quantitative data are presented as mean ± SEM. ***P < 0.001 compared with the NC mimic group. d CORO1C expression in NSCLC tissues and corresponding normal mucosa tissues were detected by q-RT-PCR. e CORO1C expression in a panel of human lung cell lines and human lung epithelia BEA-2B cells was evaluated by q-RT-PCR. CORO1C expression in BEA-2B was set as 100%. Quantitative data are presented as the mean ± SEM. * **P < 0.001 compared with BEA-2B cells

Article Snippet: Antibodies against vimentin (D21H3), E-cadherin (24E10), N-cadherin (D4R1H), ZO-1(D6L1E) and Snail (C15D3), CORO1C (D6K5B) and β-actin (13E5) were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Sequencing, Binding Assay, Expressing, Western Blot, Control, Luciferase, Mutagenesis, Activity Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Cellular & Molecular Biology Letters

Article Title: MiR-206 may suppress non-small lung cancer metastasis by targeting CORO1C

doi: 10.1186/s11658-020-00216-x

Figure Lengend Snippet: CORO1C overexpression attenuates miR-206-mediated inhibitory effect on A549 cells. a CORO1C vector transfection increased CORO1C expression. The cells were treated with CORO1C vector or NC vector for 24 h, then the cells were collected for q-RT-PCR assay. b CORO1C vector transfection decreases miR-206-mediated inhibition effect on A549 cell proliferation. The cells were transfected with NC vector and CORO1C vector, and then treated with miR-206 mimic for 24 h. Then the cells were subjected to MTT assay. c and d ) CORO1C overexpression rescues miR-206-induced effect on the horizontal migration of A549 cells. A549 cells were transfected with NC vector or CORO1C vector, and then applied for wound healing assay stimulated with miR-206 mimic. Representative images (100× magnification) and the quantitative data are shown in c and d , respectively. ( e and f ) CORO1C vector attenuates miR-206-mediated inhibitory effect on the vertical migration and invasion of A549 cells. A549 cells were transfected with NC vector or CORO1C vector, and then subjected to transwell migration and transwell invasion assays. Representative images (100 × magnification) and the quantitative data are shown in e and f , representatively. g CORO1C vector rescues miR-206-induced inhibitory effect of EMT markers. The cells were transfected with CORO1C vector or NC vector. After 24 h the cells were collected and subjected to western blotting assay. Quantitative data are presented as mean ± SEM. *P < 0.05 and * **P < 0.001 compared with the NC vector group, # P < 0.05 and ## P < 0.01 compared with the NC vector+ NC mimic group

Article Snippet: Antibodies against vimentin (D21H3), E-cadherin (24E10), N-cadherin (D4R1H), ZO-1(D6L1E) and Snail (C15D3), CORO1C (D6K5B) and β-actin (13E5) were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Over Expression, Plasmid Preparation, Transfection, Expressing, Reverse Transcription Polymerase Chain Reaction, Inhibition, MTT Assay, Migration, Wound Healing Assay, Western Blot

Journal: Oncology reports

Article Title: microRNA‑196a‑3p inhibits cell proliferation and promotes cell apoptosis by targeting ADP ribosylation factor 4 in diffuse large B‑cell lymphoma.

doi: 10.3892/or.2020.7901

Figure Lengend Snippet: Figure 4. Prediction and identification of miR‑196a‑3p target genes in DLBCL cells. (A) miRNA targets were predicted using TargetScan, microRNA, miRDB and DIANA‑microT. (B) The protein levels of 4 miR‑196a‑3p candidate target genes were analyzed by western blot analysis. Up, protein bands; down, grey value. (C) mRNA levels of 4 miR‑196a‑3p candidate target genes were analyzed by RT‑qPCR. (D) 293T cells were transiently transfected with Renilla luciferase reporter vectors containing either wild‑type or mutant ARF4 3'‑UTR seed sequence, and luciferase activity was measured following co‑transfection with miR‑196a‑3p or NC. *P<0.05, **P<0.01 and ***P<0.001. DLBCL, diffuse large B‑cell lymphoma; miRNA, microRNA; ARF4, ADP ribosylation factor 4; 3'‑UTR, 3'‑untranslated region; NC, negative control.

Article Snippet: Following endogenous peroxidase blocking and immunostaining with antibodies, the sections were incubated with the following biotinylated anti-rabbit primary antibodies: ARF4, ZNF280B, CORO1C or NRP2 (dilution 1:200; ProteinTech Group, Inc.).

Techniques: Western Blot, Transfection, Luciferase, Mutagenesis, Sequencing, Activity Assay, Negative Control

Journal: Oncology reports

Article Title: microRNA‑196a‑3p inhibits cell proliferation and promotes cell apoptosis by targeting ADP ribosylation factor 4 in diffuse large B‑cell lymphoma.

doi: 10.3892/or.2020.7901

Figure Lengend Snippet: Figure 5. Role of ARF4 in the regulation of proliferation, apoptosis and cell cycle in DLBCL cells. Farage and OCI‑LY3 cells were transfected with ARF4 siRNA or a scrambled sequence as the negative control. (A) Cell cycle analysis was performed by PI‑staining flow cytometry. (B) Cell viability was determined by the EdU staining assay. (C) Apoptosis of cells was determined by Annexin V and PI staining and flow cytometry. (D) DLBCL cell proliferation was detected by CCK‑8 assay. *P<0.05, **P<0.01 and ***P<0.001. OD, optical density; ARF4, ADP ribosylation factor 4; siARF4, ARF4 siRNA; DLBCL, diffuse large B‑cell lymphoma; siRNA, small interfering RNA; PI, propidium iodide; CCK‑8, Cell Counting Kit‑8.

Article Snippet: Following endogenous peroxidase blocking and immunostaining with antibodies, the sections were incubated with the following biotinylated anti-rabbit primary antibodies: ARF4, ZNF280B, CORO1C or NRP2 (dilution 1:200; ProteinTech Group, Inc.).

Techniques: Transfection, Sequencing, Negative Control, Cell Cycle Assay, Flow Cytometry, Staining, CCK-8 Assay, Small Interfering RNA

Journal: Oncology reports

Article Title: microRNA‑196a‑3p inhibits cell proliferation and promotes cell apoptosis by targeting ADP ribosylation factor 4 in diffuse large B‑cell lymphoma.

doi: 10.3892/or.2020.7901

Figure Lengend Snippet: Figure 6. Expression of miR‑196a‑3p and its candidate target genes in DLBCL as compared with RLH clinical specimens. (A) Representative immuno- histochemical staining for ARF4, ZNF280B, CORO1C and NRP2. (B) Protein expression was compared between DLBCL and RLH specimens using the IRS value. (C) The expression of miR‑196a‑3p was compared in two different groups based on the ARF4 protein level. DLBCL, diffuse large B‑cell lym- phoma; RLH, reactive lymph node hyperplasia; ARF4, ADP ribosylation factor 4; ZNF280B, Zinc finger protein; CORO1C, coronin‑1C; NRP2, neuropilin‑2; IRS, immunoreactive score.

Article Snippet: Following endogenous peroxidase blocking and immunostaining with antibodies, the sections were incubated with the following biotinylated anti-rabbit primary antibodies: ARF4, ZNF280B, CORO1C or NRP2 (dilution 1:200; ProteinTech Group, Inc.).

Techniques: Expressing, Staining