Journal: Viruses

Article Title: The MAPK/ERK Pathway and the Role of DUSP1 in JCPyV Infection of Primary Astrocytes

doi: 10.3390/v13091834

Figure Lengend Snippet: ERK knockdown significantly reduced JCPyV infection in NHAs. NHAs, SVGAs, and NHA-Ts were transfected with ERK1/2 or EGFR (CTL) siRNA. At 72 h post-transfection, cells were fixed for analysis of ERK protein expression ( A , B ) or infected ( C ). ( A ) ERK1/2 protein expression was determined by in-cell western (ICW), staining for Total ERK1/2 (green) or CellTag (red). ( B ) Percentage of protein knockdown was measured by ICW signal intensity values per [(ERK1/2)/Cell Tag × 100% = % response] within each ICW analysis using LI-COR software. Error bars indicate SD. Student’s t -test was used to determine statistical significance comparing EGFR (CTL) siRNA-treated cells to ERK1/2 siRNA-treated cells, for each cell type. *, p < 0.01. ( C ) Following 3 days post-transfection, cells were infected with JCPyV (MOI = 1.0 FFU/cell) at 37 °C for 1 h. Cells were incubated in complete media at 37 °C for 48 h and then fixed and stained by indirect immunofluorescence. Infectivity following ERK1/2 knockdown ( C ) was determined by counting the number of JCPyV T Ag-positive nuclei divided by the number of DAPI-positive nuclei for five × 20 fields of view for triplicate samples. Error bars indicate SD. Student’s t test was used to determine statistical significance comparing EGFR (CTL) siRNA-treated cells to ERK1/2 siRNA-treated cells for each cell type ( C ). * p < 0.01.

Article Snippet: NHAs and NHA-Ts were plated for ~70% confluency, while SVGAs were plated to ~50% confluency in 96 well plates and transfected with siRNAs specific for ERK1/2, epidermal growth factor receptor (EGFR) control (Cell Signaling Technology, Danvers, MA, USA), or DUSP1 (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Knockdown, Infection, Transfection, Expressing, In-Cell ELISA, Staining, Software, Incubation, Immunofluorescence

Journal: Viruses

Article Title: The MAPK/ERK Pathway and the Role of DUSP1 in JCPyV Infection of Primary Astrocytes

doi: 10.3390/v13091834

Figure Lengend Snippet: DUSP1 was required for JCPyV infection in NHAs, regardless of pERK expression. NHAs, SVGAs, and NHA-Ts were transfected with DUSP1 or EGFR (CTL) siRNA. At 24 h post-transfection, cells were fixed, imaged, and quantified for protein knockdown ( A , B ) or infected ( C ). ( A ) DUSP1 protein expression was determined by ICW, staining for total DUSP1 (green) or CellTag (red). ( B ) Quantification of DUSP1 protein expression was measured by ICW signal intensity values per [(DUSP1)/Cell Tag × 100% = % response] within each ICW analysis in LI-COR software. Cells treated with DUSP1 siRNA were normalized to the respective EGFR (CTL) siRNA (dashed line). Box and whisker plots represent the distribution of 9 samples, with the median denoted by the black line and whiskers representing values 1.0 times the distance of the inter-quartile range. Outliers are represented by black diamonds. A pairwise Wilcoxon rank-sum test, along with the Bonferroni adjustment, was used to compare CTL and DUSP1 siRNA-treated cells across each cell type. *, p < 0.01. Data are representative of three independent experiments performed in triplicate. ( C ) At 24 h post-transfection, cells were infected with JCPyV (MOI = 1.0 FFU/cell) at 37 °C for 1 h. Cells were incubated in complete media for 48 h then fixed and stained by indirect immunofluorescence. Percent infection following DUSP1 knockdown was determined by quantifying the number of JCPyV T Ag-positive nuclei divided by the number of DAPI-positive nuclei for five × 20 fields of view for triplicate samples. Error bars indicate SD. Student’s t test was used to determine statistical significance comparing EGFR (CTL) siRNA-treated cells to DUSP1 siRNA-treated cells, for each cell type. *, p < 0.01. ( D ) Quantification of pERK expression following DUSP1 siRNA at 24 h post-transfection was measured by ICW signal intensity values per [(pERK)/Cell Tag × 100% = % response] within each ICW analysis in LI-COR software. Cells treated with DUSP1 siRNA were normalized to the EGFR (CTL) siRNA (dashed line). Box and whisker plots represent the distribution of 9 samples, with the median denoted by the black line and whiskers representing values 1.0 times the distance of the inter-quartile range. Outliers are represented by black diamonds. A Wilcoxon rank-sum exact test was used to compare CTL and DUSP1 siRNA-treated cells. *, p < 0.05.

Article Snippet: NHAs and NHA-Ts were plated for ~70% confluency, while SVGAs were plated to ~50% confluency in 96 well plates and transfected with siRNAs specific for ERK1/2, epidermal growth factor receptor (EGFR) control (Cell Signaling Technology, Danvers, MA, USA), or DUSP1 (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Infection, Expressing, Transfection, Knockdown, Staining, Software, Whisker Assay, Incubation, Immunofluorescence

Journal: Scientific Reports

Article Title: Leucyl-tRNA synthetase deficiency systemically induces excessive autophagy in zebrafish

doi: 10.1038/s41598-021-87879-4

Figure Lengend Snippet: Inhibition of autophagy prevents abnormal development and improves survival in larsb- knockout larvae. (A) Morphology of larsb + / + and larsb −/− embryos injected with either control MO or atg5-MO (72 h post fertilization (hpf)). Scale bars: 500 µm. (B) Western blot analysis of Lc3b protein expression at 72 hpf for wild-type embryos injected with either control MO or atg5-MO. β-actin levels served as the loading control. (C) Morphological abnormality at 72 hpf in the livers of larsb −/− larvae under Tg[ fabp10 :mcherry] background injected with either control MO or atg5-MO. Scale bars: 200 μm. (D) Quantification of liver size in larsb −/− larvae under Tg[ fabp10 :mcherry] background (72 hpf). Liver sizes were evaluated using ImageJ software version 1.52a ( https://imagej.nih.gov/ij/ ). n = 4 fish/group. Error bars indicate SEM. Student’s t-test; ***P < 0.001. (E) Western blot analysis of Lc3b protein expression at 72 hpf for wild-type embryos treated with DMSO or bafilomycin A1. β-actin levels served as the loading control. (F) Morphological abnormality at 72 hpf in the livers of larsb −/− larvae under Tg[ fabp10 :mcherry] background treated with DMSO or bafilomycin A1. Scale bars: 200 μm. (G) Quantification of liver size in larsb −/− larvae under Tg[ fabp10 :mcherry] background (72 hpf). Liver sizes were evaluated using ImageJ software version 1.52a ( https://imagej.nih.gov/ij/ ). n = 10 fish/group. Error bars indicate SEM. Student’s t-test; *P < 0.05. (H) Kaplan–Meier survival curve of larsb + / + (n = 23) and larsb −/− (n = 15) larvae treated with DMSO and larsb + / + (n = 11) and larsb −/− larvae (n = 21) treated with bafilomycin A1. Statistics were calculated and the figure was produced in GraphPad software version 8 ( https://www.graphpad.com/scientific-software/prism/ ). Larsb: leucyl-tRNA synthetase b, MO: morpholino, n.s.: non-significant, DMSO: dimethyl sulfoxide, Dpf: days post fertilization.

Article Snippet: Western blotting was performed with antibodies against Lars (#13868; Cell Signaling Technology, Beverly, MA, USA), p62 (PM045; Medical & Biological Laboratories, Nagoya, Japan), LC3B (PM036; Medical & Biological Laboratories), ATG5 (NB110-53818; Novus Biologicals, Littleton, CO, USA), β-actin (A3854; Sigma-Aldrich, St. Louis, MO, USA), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (G9295; Sigma-Aldrich).

Techniques: Inhibition, Knock-Out, Injection, Control, Western Blot, Expressing, Software, Produced

Journal: Cell reports

Article Title: Glioblastoma Cell Resistance to EGFR and MET Inhibition Can Be Overcome via Blockade of FGFR-SPRY2 Bypass Signaling

doi: 10.1016/j.celrep.2020.02.014

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Phospho-EGF Receptor (Tyr1068) (D7A5) XP Rabbit mAb , Cell Signaling Technology , RRID:AB_2096270.

Techniques: Virus, Subcloning, Recombinant, Chromatin Immunoprecipitation, Ab Array, Enzyme-linked Immunosorbent Assay, Proliferation Assay, Bicinchoninic Acid Protein Assay, Stable Transfection, Expressing, shRNA, Sequencing, Software

Journal: Aging (Albany NY)

Article Title: Restoration of aged hematopoietic cells by their young counterparts through instructive microvesicles release

doi: 10.18632/aging.203689

Figure Lengend Snippet: Key Resources Table.

Article Snippet: Anti-Human Cyclin D1 (WB) , Cell Signaling , Cat #2978.

Techniques: Recombinant, Saline, Protease Inhibitor, SYBR Green Assay, Isolation, Cytotoxicity Assay, Viability Assay, Software, Expressing, RNA Sequencing, Sequencing, Real-time Polymerase Chain Reaction

Journal: The EMBO Journal

Article Title: Ephrin A1 functions as a ligand of EGFR to promote EMT and metastasis in gastric cancer

doi: 10.1038/s44318-025-00363-x

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Phospho-EGF Receptor (Tyr1068) Rabbit mAb , CST , Cat# 3777.

Techniques: Recombinant, Sequencing, Software

Journal: Cell Death & Disease

Article Title: HER3 promotes triple-negative breast cancer progression by upregulating PHF8 via miR-34b-5p-dependent mechanism

doi: 10.1038/s41419-025-08115-9

Figure Lengend Snippet: A Volcano plot for RNA-Seq analyses of differentially expressed genes (DEGs) in HCC1806 cells with or without HER3 depletion. Each point represented the average value of one transcript in three replicates. The difference was considered significant for adjusted p < 0.05 (red for upregulated genes, blue for downregulated genes, and gray for non-significant genes). The indicated genes were involved in chromatin organization and histone modification. B Gene Ontology (GO) enrichment analysis of the DEGs. Top GO terms were represented by gene count >50, with FDR < 0.05 in Molecular Function (MF), Cellular Component (CC) and Biological Process (BP). C KEGG pathway analysis of the DEGs. D , E HCC1806, MDA-MB-231 (MDA-231), and MDA-MB-468 (MDA-468) cells were infected with lentivirus containing either control shRNA (shControl) or specific shRNA against HER3 (sh HER3 ) for 48 h. Cell cycle distribution was analyzed with flow cytometry assays ( D ). Western blot analyses were performed to examine p-HER3, HER3, p-Akt, Akt, E2F1, Cyclin D1, p27 kip1 , p21 waf1 , and β-actin ( E ). F The heat map showed the top ten (down- or up-regulated) genes involved in chromatin organization and histone modification. Analysis of TCGA ( G ) and CCLE ( H ) datasets revealed a significantly positive correlation between HER3 and PHF8 expression in breast cancer specimens and cell lines, respectively.

Article Snippet: Primary antibodies used in western blot assays included HER3 (cat#12708, 1:1000), p-HER3 (Y1289) (cat#4791, 1:1000), p-Akt (S473) (cat#9271, 1:1000), Akt (cat#9272, 1:1000), PHF8 (cat#93801, 1:1000), p27 kip1 (cat#3686, 1:1000), p21 (cat#2947, 1:1000), Cyclin D1 (cat# 2922, 1:1000), E2F1 (cat# 3742, 1:1000) from Cell Signaling Technology (Beverly, MA, USA), and β-actin (A3853, 1:10,000) from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: RNA Sequencing, Modification, Infection, Control, shRNA, Flow Cytometry, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: HER3 promotes triple-negative breast cancer progression by upregulating PHF8 via miR-34b-5p-dependent mechanism

doi: 10.1038/s41419-025-08115-9

Figure Lengend Snippet: TNBC cell lines (HCC1806, HCC70, HCC1937, and MDA-MB-468) were transiently infected with lentivirus containing either control shRNA (sc) or specific shRNAs targeting PHF8 (sh1 and sh2). A Cells were seeded onto 6-well plates for colony formation assays. Representative images of the cell colonies were captured using a digital camera after crystal violet staining (left panel), and the colony numbers were quantified using ImageJ software (right panel). Error bars represent the standard deviation (SD). *, p < 0.05; **, p < 0.01; #, p < 0.005. B Cell cycle progression was examined by flow cytometry analysis. C Western blot was assays were conducted to detect PHF8, E2F1, Cyclin D1, p27 kip1 , p21 waf1 , and β-actin. D , RT-qPCR was performed to measure PHF8 and p27 kip1 mRNA expression. E , F HCC1806 and HCC1937 cells were infected with lentivirus containing either control shRNA (-) or HER3 -specific shRNA (+). After 48 h, cells were collected and subjected to western blot analysis of HER3, PHF8, p27 kip1 , and β-actin ( E ). RT-qPCR analysis was performed to quantify the mRNA expression levels of HER3, PHF8, and p27 kip1 ( F ). G HCC1806 and HCC1937 cells with ectopic expression of HER3 were subjected to western blot analysis of HER3, PHF8, p27 kip1 , and β-actin. H TNBC cells infected with lentivirus containing either control shRNA (-) or PHF8 -specific shRNA (+) were stimulated with HRG-β1 (25 ng/ml) for 24 h. Cells were collected and examined by western blot analysis of p-HER3, PHF8, p27 kip1 , and β-actin.

Article Snippet: Primary antibodies used in western blot assays included HER3 (cat#12708, 1:1000), p-HER3 (Y1289) (cat#4791, 1:1000), p-Akt (S473) (cat#9271, 1:1000), Akt (cat#9272, 1:1000), PHF8 (cat#93801, 1:1000), p27 kip1 (cat#3686, 1:1000), p21 (cat#2947, 1:1000), Cyclin D1 (cat# 2922, 1:1000), E2F1 (cat# 3742, 1:1000) from Cell Signaling Technology (Beverly, MA, USA), and β-actin (A3853, 1:10,000) from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Infection, Control, shRNA, Staining, Software, Standard Deviation, Flow Cytometry, Western Blot, Quantitative RT-PCR, Expressing

Journal: STAR Protocols

Article Title: Protocol for evaluating the effects of large gradient high magnetic fields on osteocyte function

doi: 10.1016/j.xpro.2024.103186

Figure Lengend Snippet: Protein expression levels of Wnt signaling pathway (Wnt1, LRP6, β-catenin, and SOST) Data shown as mean ± SD. n = 3. ∗ p < 0.05, ∗∗ p < 0.01; vs. control. ## p < 0.01, ### p < 0.001; 12 T upward vs. 12 T downward.

Article Snippet: WNT1 polyclonal antibody (1:1,000) , Proteintech , RRID: AB_2881013 ; Cat# 27935-1-AP.

Techniques: Expressing, Control

Journal: STAR Protocols

Article Title: Protocol for evaluating the effects of large gradient high magnetic fields on osteocyte function

doi: 10.1016/j.xpro.2024.103186

Figure Lengend Snippet:

Article Snippet: WNT1 polyclonal antibody (1:1,000) , Proteintech , RRID: AB_2881013 ; Cat# 27935-1-AP.

Techniques: Recombinant, Software, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Cell Culture, Western Blot

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Cyclin D1 is a major target of miR-206 in cell differentiation and transformation.

doi: 10.4161/cc.26674

Figure Lengend Snippet: Figure 1. miR-206 targets cyclin D1. (A) Sequence alignment between miR-206 and the 3′UTRs of cyclin D1 from different species. In brackets the 3′UTR size. (B) Diagram of the luciferase reporter construct with the putative miR-206 binding site (WT 3′UTR) and mutations (3′UTR MUT). (C) Relative lucif- erase activity was measured in HeLa cells after transfection of reporter constructs along with pSP65-U1 (CTR) or pSP65–206 (miR-206). Relative Firefly luciferase values were determined by a ratio of Firefly to Renilla luciferase with the control set to 1.00. Values are the means ± SD of 3 separate experi- ments. **A Student t test performed between control and miR-206 transfected cells yielded P values < 0.01.

Article Snippet: The mouse monoclonal antibody 72–13G (Santa Cruz Biotechnology) was used to stain cyclin D1-positive cells.

Techniques: Sequencing, Luciferase, Construct, Binding Assay, Activity Assay, Transfection, Control

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Cyclin D1 is a major target of miR-206 in cell differentiation and transformation.

doi: 10.4161/cc.26674

Figure Lengend Snippet: Figure 2. Expression kinetics of miR-206 and cyclin D1 in differentiating C2C12 cells. C2C12 myoblasts were seeded in GM at 1.5 × 104/cm2. Cells were shifted in DM 24 h after plating and left to differentiate for further 72 h. (A) Northern blot analysis of miR-206 expression in C2C12 cells after 24 h in GM (0) and at different time points upon shift to DM. (B) Western blot analysis of cyclin D1 and MyHC expression in C2C12 cells cultured as in (A). Equal RNA and protein loading was confirmed by detecting, snRNA U2 and β-tubulin, respectively. (C) MyHC immunofluorescence staining (green) of C2C12 cells after 24 h in GM (DM 0 h) and after 72 h in DM (DM 72 h). Nuclei were counterstained in blue (DAPI) and individual pictures of the same field, taken with a DC camera, were merged using a LEICA Microsystems Imaging Equipment. Bar = 20 μm.

Article Snippet: The mouse monoclonal antibody 72–13G (Santa Cruz Biotechnology) was used to stain cyclin D1-positive cells.

Techniques: Expressing, Northern Blot, Western Blot, Cell Culture, Immunofluorescence, Staining, Imaging

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Cyclin D1 is a major target of miR-206 in cell differentiation and transformation.

doi: 10.4161/cc.26674

Figure Lengend Snippet: Figure 3. miR-206 controls cyclin D1 accumulation in C2C12 cells. C2C12 myoblasts were seeded in GM at 2.5 × 103/cm2. Cells were transfected 24 h after plaiting. (A) Northern blot analysis of miR-206 expression (upper) and western blot analysis of cyclin D1 expression (lower) in C2C12 cells 48 h after transfection with a control vector (CTR) or with a miR-206 expression vector (miR-206). Cells were kept in GM throughout the experiment. (B) The effect of miR-206 overexpression on C2C12 cell proliferation and differentiation was evaluated 48 h after transfection by 1 h BrdU incorporation and MyHC staining, respectively. Results are represented relative to the BrdU+ nuclei or nuclei in MyHC+ cells in CTR (set to 1.00), as individually assessed in each independent experiment. Values are the means ± SD of 3 separate experiments. *A Student t test performed between control and miR-206 transfected cells yielded P values < 0.05. (C) Immunofluorescence staining of cyclin D1 (pink) and MyHC (green) 48 h after transfection. Nuclei were counterstained in blue with DAPI. Individual pictures of the same field, taken with a DC camera, were merged using a LEICA Microsystems Imaging Equipment. To obtain cyclin D1 images, before merging, individual pictures were pseudocol- ored using a LEICA Microsystems Imaging software. Bar = 10 μm. (D) C2C12 myoblasts were seeded at low (LD) and high (HD) density in GM. Cells were shifted to DM the day after plating and analyzed after further 3 d. The panels show a northern blot analysis of miR-206 expression (left panel) and a western blot analysis of cyclin D1 and differentiation associ- ated marker expression (right panel) after 24 h in GM and 72 h after shift- ing to DM. Equal RNA and protein loading was confirmed by detecting, snRNA U2 and β-tubulin, respectively.

Article Snippet: The mouse monoclonal antibody 72–13G (Santa Cruz Biotechnology) was used to stain cyclin D1-positive cells.

Techniques: Transfection, Northern Blot, Expressing, Western Blot, Control, Plasmid Preparation, Over Expression, BrdU Incorporation Assay, Staining, Immunofluorescence, Imaging, Software, Marker

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Cyclin D1 is a major target of miR-206 in cell differentiation and transformation.

doi: 10.4161/cc.26674

Figure Lengend Snippet: Figure 4. Inhibition of miR-206 rescues cyclin D1 in myotubes (A) Experimental scheme. C2C12 myoblasts were induced to differ- entiate in DM in the presence of AraC. After 3 d, AraC was washed out and cells left to recover in DM for further 24 h. Finally, pure myotubes were transfected with LNA against miR-206 and analyzed 48 h later. (B) Northern blot analysis of miR-206 and miR-1 expression (left panel) and western blot analysis of cyclin D1 expression (right panel) in pure myotubes transfected with a control LNA (LNA C) or anti-miR-206 LNA (LNA 206). Cyclin D1 expression in proliferating myoblasts is also shown (GM). Equal RNA and protein loading was confirmed by detecting, snRNA U2 and β-tubulin, respectively. (C) Double immunofluorescence staining of MyHC and cyclin D1 of pure myotubes transfected with a control LNA (LNA C) or anti-miR-206 LNA (LNA 206). Individual pictures of the same field, taken with a DC camera, were merged using a LEICA Microsystems Imaging Equipment. Bar = 10 μm.

Article Snippet: The mouse monoclonal antibody 72–13G (Santa Cruz Biotechnology) was used to stain cyclin D1-positive cells.

Techniques: Inhibition, Transfection, Northern Blot, Expressing, Western Blot, Control, Double Immunofluorescence Staining, Imaging

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Cyclin D1 is a major target of miR-206 in cell differentiation and transformation.

doi: 10.4161/cc.26674

Figure Lengend Snippet: Figure 5. miR-206 inhibits cell proliferation in Ras-transformed fibro- blasts. (A) Expression levels of cyclin D1 in NIH3T3(Ras) cells as compared with NIH3T3(BN) cells. (B) Real-time PCR analysis of miR-206 expres- sion in NIH3T3(Ras) cells. Results are shown relative to untransformed NIH3T3(BN) cells set to value 1.00. Each sample was analyzed in tripli- cate, and values are the means ± SD of 3 independent experiments. **A Student t test performed between untransformed and transformed cells yielded P values < 0.01. (C) NIH3T3(Ras) cells were transfected with a control vector (CTR) or with a miR-206 expression vector (miR-206) and analyzed 24 h later. Upper, northern blot analysis of miR-206 expression; lower, western blot analysis of cyclin D1 expression. (D) Effect of miR-206 forced expression on cell proliferation as determined by 1 h BrdU incor- poration. Data are reported relative to BrdU+ nuclei in CTR (set to 1.00), as individually assessed in each independent experiment. Values are the means ± SD of 3 separate experiments. *A Student t test performed between control and miR-206 transfected cells yielded P values < 0.05. Equal RNA and protein loading was confirmed by detecting, snRNA U2, and β-tubulin, respectively.

Article Snippet: The mouse monoclonal antibody 72–13G (Santa Cruz Biotechnology) was used to stain cyclin D1-positive cells.

Techniques: Transformation Assay, Expressing, Real-time Polymerase Chain Reaction, Transfection, Control, Plasmid Preparation, Northern Blot, Western Blot

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Cyclin D1 is a major target of miR-206 in cell differentiation and transformation.

doi: 10.4161/cc.26674

Figure Lengend Snippet: Figure 6. Relationship between miR-206 downregulation and cyclin D1 expression in NSCLCs. (A) Northern blot analysis of miR-206 in different murine tissues. snRNA U2 levels were used as a loading control. (B) Real-time PCR analysis of miR-206 expression in human NSCLC tissues. Results are shown relative to the matched normal lung tissues set to value 1.00. Each sample was analyzed in triplicate, and values are the means ± SD of three independent experiments. **A Student t test performed between normal and tumor tissues yielded P values < 0.01. (C) Western blot analysis of cyclin D1 expression in normal and neoplastic lung tissues. Equal protein loading was confirmed by detecting actin. n, normal tissue; t = tumor tissue

Article Snippet: The mouse monoclonal antibody 72–13G (Santa Cruz Biotechnology) was used to stain cyclin D1-positive cells.

Techniques: Expressing, Northern Blot, Control, Real-time Polymerase Chain Reaction, Western Blot

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Cyclin D1 is a major target of miR-206 in cell differentiation and transformation.

doi: 10.4161/cc.26674

Figure Lengend Snippet: Figure 7. miR-206 inhibits cancer cell proliferation through repression of cyclin D1. (A) A549 and HeLa cells were transfected with a control vec- tor (CTR) or with a miR-206 expression vector (miR-206) and analyzed 72 h later. Top panel: northern blot analysis of miR-206 expression; lower panel, western blot analysis of cyclin D1 expression. Equal RNA and protein loading was confirmed by detecting, snRNA U2 and β-tubulin, respectively. (B) Effect of miR-206 forced expression on cell prolifera- tion as determined by 1 h BrdU incorporation and immunofluorescence staining. Data are reported relative to BrdU+ nuclei in CTR (set to 1.00), as individually assessed in each independent experiment. Values are the means ± SD of 3 separate experiments. *A Student t test performed between control and miR-206 transfected cells yielded P values <0.05.

Article Snippet: The mouse monoclonal antibody 72–13G (Santa Cruz Biotechnology) was used to stain cyclin D1-positive cells.

Techniques: Transfection, Control, Expressing, Plasmid Preparation, Northern Blot, Western Blot, BrdU Incorporation Assay, Immunofluorescence, Staining

Journal: iScience

Article Title: SGMS1 facilitates osteogenic differentiation of MSCs and strengthens osteogenesis-angiogenesis coupling by modulating Cer/PP2A/Akt pathway

doi: 10.1016/j.isci.2024.109358

Figure Lengend Snippet:

Article Snippet: Akt Mouse monoclonal antibody , Proteintech , 60203-2-Ig; RRID:AB_2919902.

Techniques: Recombinant, Virus, Modification, Reverse Transcription, SYBR Green Assay, IP Phosphatase Assay, Software