Journal: PeerJ

Article Title: CTHRC1 promotes anaplastic thyroid cancer progression by upregulating the proliferation, migration, and invasion of tumor cells

doi: 10.7717/peerj.15458

Figure Lengend Snippet: Western blotting was used to analyze the β-catenin, Cyclin D1, and c-myc expression in CTHRC1 knockdown and control group. ***P < 0.001 .

Article Snippet: After being blocked with 5% non-fat milk for 2 h, the membranes were probed with primary antibodies against CTHRC1 (1:1000, ab256458; Abcam, Cambridge, UK), β-catenin (1:1000, A00004; Boster Bio, Wuhan, China), c-myc (1:500, BA1284-2; Boster Bio, Wuhan, China), Cyclin D1 (1:1000, BA0770-2; Boster Bio, Wuhan, China), E-Cadherin (1:1000, ab235682; Abcam, Cambridge, UK), Vimentin (1:1000, #5741; Cell Signaling Technology, Danvers, MA, USA), Snail (1:2000, A5243; ABclonal Science Inc., Woburn, MA, USA), β-actin (1:5000, AC026; ABclonal, Woburn, MA, USA), GAPDH (1:10000, TA802519BM; Origene, Rockville, MD, USA), overnight at 4 °C, after which they were subsequently incubated with the secondary antibodies (1:5000, ab150077/ab150113; Abcam, Cambridge, UK) for 40 min, at 25 °C.

Techniques: Western Blot, Expressing, Knockdown, Control

Journal: BMC cancer

Article Title: NR1D1 suppressed the growth of ovarian cancer by abrogating the JAK/STAT3 signaling pathway.

doi: 10.1186/s12885-021-08597-8

Figure Lengend Snippet: Fig. 2 NR1D1 induced cell cycle arrest and apoptosis in ovarian cancer cells. A Cell cycle of NR1D1 over-expressed cells was determined by flow cytometry. B The levels of cyclinD and cyclinE were determined by western blot. C Flow cytometry was performed to determine the cell cycle of NR1D1 silenced cells. D Western blot was performed to determine the levels of cyclinD and cyclinE. E Apoptosis of NR1D1 over-expressed cells was determined by flow cytometry. F After transfection with NR1D1 over-expression plasmid, the levels of activated caspase-3 and caspase-9 were determined. The results are presented as mean + SD

Article Snippet: Following blocking with 5% bovine serum albumin, the membranes were incubated with antibodies against NR1D1 (1:1000; Abclonal, Wuhan, China), cyclinD (1:1000; ABclonal), cyclinE (1:1000; Proteintech, Wuhan, China), SOCS3 (1:1000; ABclonal), JAK-1 (1:1000; Affinity, Changzhou, China), p-JAK1 (Tyr 1034/Tyr 1035; 1:1000; Affinity), JAK2 (1:500; Affinity), p-JAK2 (Tyr 1007/Tyr 1008, 1:1000; Affinity), STAT3 (1:500; Affinity), p-STAT3 (Tyr 705, 1:500; Affinity), β-actin (1:2000; Proteintech) at 4 °C overnight.

Techniques: Flow Cytometry, Western Blot, Transfection, Over Expression, Plasmid Preparation

Journal: ACS Applied Materials & Interfaces

Article Title: Dual-Targeted Biomimetic Nanoparticles for Enhanced Delivery of Polyphyllin B Synergistically Induce Ferroptosis and Immunogenic Cell Death in Gastric Cancer

doi: 10.1021/acsami.5c13198

Figure Lengend Snippet: Ma/HA@NPs-PB promoted the apoptosis of gastric cancer cells. (A, B) Statistical analysis of the cell cycle of HGC27 and SGC7901 cells after 72 h of incubation with PB, HA@NPs-PB or Ma/HA@NPs-PB. (C–G) Expression of CDK4, CDK6 and Cyclin D1 proteins in each group after coculture with HGC27 and SGC7901 cells and statistical analysis. (H–L) Immunofluorescence images and statistical analysis of Ki67 and Bcl-2 after coculture with HGC27 (left) and SGC7901 (right) cells in each group. Ki67 (green), Bcl-2 (red), and DAPI (blue) are shown. (M, N) Statistical analysis of the apoptosis of HGC27 and SGC7901 cells after incubation with PB, HA@NPs-PB or Ma/HA@NPs-PB for 3 and 5 days. The data are expressed as the means ± SD ( n = 3). ns indicates no statistical significance. * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001.

Article Snippet: Then, CDK4 (Cat. 83994-3-RR, Proteintech, China), CDK6 (Cat. 14052-1-AP, Proteintech, China), CyclinD1 (Cat. 26939-1-AP, Proteintech, China), GPX4 (Cat. 30388-1-AP, Proteintech, China), HER2 (Cat. 18299-1-AP, Proteintech, China), CD44 (Cat. 15675-1-AP, Proteintech, China), galectin-3 (Cat. 14979-1-AP, Proteintech, China), CD47 (Cat. 20305-1-AP, Proteintech, China), HMGB1 (Cat. 18256, Abcam, US) and CRT (Cat. 92516, Abcam, US) were incubated overnight at 4 °C.

Techniques: Incubation, Expressing, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Cyclin D1/D2–CDK4 Drives Cell Migration by Orchestrating Cytoskeletal Dynamics Through a TGFβ–FAK–Rac1 Axis

doi: 10.3390/ijms27031228

Figure Lengend Snippet: Co-localization of CDK4 with cyclin D1/D2 at the lamellipodia region. ( A ) Subcellular localization of CDK4 and CDK6 in wild-type HeLa cells. Cells were stained for F-actin (FITC-phalloidin, green), CDK4 or CDK6 (purple), and nuclei (DAPI, blue). Scale bar, 10 μm. ( B ) The line-scan profile shows the fluorescence intensity across the membrane ruffles, cytoplasm, and nucleus along the line drawn on the CDK4 image in panel ( A ). ( C ) Subcellular localization of cyclin D1, D2, and D3 (red) in wild-type HeLa cells. Scale bar, 10 μm. ( D , E ) The line-scan profile shows the fluorescence intensity across the membrane ruffles, cytoplasm, and nucleus along the line drawn on the cyclin D1/D2 images in panel ( C ). ( F ) Immunofluorescence staining showing co-localization of CDK4 (purple) and cyclin D1 (red) at the lamellipodia region. ‘Merge’ indicates the original merged image; ‘Merge (saturation-enhanced)’ shows the merged image after saturation adjustment to better visualize the co-localization of CDK4 with cyclin D1 within lamellipodia. Scale bar, 10 μm. ( G ) The linear intensity profile along the line shown in the Merge image of panel ( F ). Pearson’s correlation coefficient (PCC) R = 0.9333. ( H ) Immunofluorescence staining showing co-localization of CDK4 (purple) and cyclin D2 (red) at the lamellipodia region. Scale bar, 10 μm. ( I ) The intensity profile along the line shown in the Merge image of panel ( H ). PCC R = 0.8727.

Article Snippet: Cyclin D1 (Proteintech, Wuhan, China, 26939-1-AP, 1:500 for immunofluorescence (IF)), Cyclin D2 (Proteintech, Wuhan, China, 10934-1-AP, 1:500 for IF), Cyclin D3 (Proteintech, Wuhan, China, 26755-1-AP, 1:500 for IF), CDK4 (Santa Cruz Biotechnology, Dallas, TX, USA, sc-23896, 1:500 for IF), CDK6 (Santa Cruz Biotechnology, Dallas, TX, USA, sc-7961, 1:500 for IF), Phospho-FAK (Tyr397) (Cell Signaling Technology, Danvers, MA, USA, 3283S, 1:1000 for immunoblotting (IB)), FAK (Cell Signaling Technology, Danvers, MA, USA, 3285S, 1:1000 for IB), Paxillin (Abcam, Waltham, MA, USA, ab32084, 1:50 for IF), GAPDH (Proteintech, Wuhan, China, 10494-1-AP, 1:5000 for IB).

Techniques: Staining, Fluorescence, Membrane, Immunofluorescence