anti clip1  (Proteintech)

Journal: Ecotoxicology and environmental safety

Article Title: Exosomal circCLIP1 regulates PM 2.5 -induced airway obstruction via targeting SEPT10 in vitro.

doi: 10.1016/j.ecoenv.2023.114750

Figure Lengend Snippet: Fig. 2. Identification of loop structure and localization of circCLIP1. A. Sanger sequencing confirmed the head-to-tail splicing of circCLIP1. B. Expression of circular and linear CLIP1 after amplified by random hexamer or Oligo (dT)18 primer was determined by RT-qPCR. C. Expression of back-spliced and canonical form of CLIP1 in cDNA and gDNA was measured by agarose gel electrophoresis assay. D. Expression of circular and linear CLIP1 with/without RNase R treatment was assessed by RT-qPCR. E. Subcellular localization of circCLIP1 was detected by fluorescence in situ hybridization. Scale bar, 50 µm. F. Expression of circCLIP1 in culture medium of PM2.5-treated HBE cells treated with DMSO or GW4869 was determined by RT-qPCR. Data were represented as means ± SD (n = 3). Statistical significance was assessed using two-tailed Student’s t test, * P < 0.05.

Article Snippet: The primary antibodies included Alix (92880, CST, USA), CD63 (ab134045, Abcam, USA), MUC5AC (ab198294, Abcam, USA), CLCA1 (ab180851, Abcam, USA), β-actin (66009–1-Ig, Proteintech, China), GAPDH (60004–1-Ig, Proteintech, China), SM-MHC (ab53219, Abcam, USA), α-SMA (19245, Cell Signaling Technology, USA), RhoA (2117, Cell Signaling Technology, USA), SEPT10 (A304–915A, ThermoFisher, USA), DKC1 (ab156877, Abcam, USA) and CLIP1 (23839–1-AP, Proteintech, China).

Techniques: Sequencing, Expressing, Amplification, Random Hexamer, Quantitative RT-PCR, Agarose Gel Electrophoresis, Fluorescence, In Situ Hybridization, Two Tailed Test

Journal: Cells

Article Title: Human Placental Endothelial Cell and Trophoblast Heterogeneity and Differentiation Revealed by Single-Cell RNA Sequencing

doi: 10.3390/cells12010087

Figure Lengend Snippet: Cell subtype identification of human placental trophoblast cells. ( A ) UMAP plot showing eight subtypes of human placental trophoblast cells. ( B ) Violin plots showing the expression levels of representative marker genes across the eight clusters. Y -axis demonstrates log scale normalized read count. ( C , D ) Heatmap and GO enriched terms of cell type differentially expressed genes among the five VCT clusters. ( E ) Cell cycle analysis of eight trophoblast clusters. ( F ) Heatmap and the clustering structure show the dissimilarity across the trophoblast subpopulations. Blue color denotes the high similarity and red color denotes the low similarity. ( G ) Violin plots showing the expression levels of a representative syncytin-related gene set ( ERVFRD-1 , MFSD2A , ERVW-1 , ASCT1 , ASCT2 ) for each trophoblast cluster. ( H ) Immunofluorescence staining for the indicated VCT-5 marker CLIP1 and trophoblast marker EGFR in 6 weeks of gestation placenta. The white arrowheads indicate EGFR + CLIP1 + cells. Scale bars, 20 μm. ( I , J ) In vitro analysis confirmed CLIP1 is induced under fusion differentiation conditions. BeWo trophoblast cells were exposed to differentiating conditions (50 μM forskolin) for 0, 6, 12, 24, or 48 h. qRT-PCR ( I ) and Western blotting ( J ) showed the expression of CLIP1 and SCT marker genes mRNA and protein levels evaluated. * p < 0.05, n = 3.

Article Snippet: The antibodies we used were: anti-CLIP1 (Abcam, ab61830, 1:1000), anti-TBX3 (Abcam, ab99302, 1:1000), anti-ERVFRD-1 (Abcam, ab230235, 1:1000), anti-GAPDH (Cell Signaling Technology, #5174, 1:2000), anti-rabbit-IgG (Cell Signaling Technology, #7074, 1:5000), and anti-mouse-IgG (Cell Signaling Technology, #7076, 1:5000).

Techniques: Expressing, Marker, Cell Cycle Assay, Immunofluorescence, Staining, In Vitro, Quantitative RT-PCR, Western Blot

Journal: The Journal of Cell Biology

Article Title: EB1 and EB3 regulate microtubule minus end organization and Golgi morphology

doi: 10.1083/jcb.201701024

Figure Lengend Snippet: Effect of EB disruption on +TIP localization. (A and C) Immunostaining for CLIP170 and CLASP1 in control and EB1/2/3mut HeLa cells expressing the indicated constructs. Enlargements of the boxed areas indicated by numbers are shown on the right. In C, cells were incubated with the GSK3 inhibitor SB415286 (20 µM) for 30 min before fixation. (B and D) Averaged intensities of staining for the indicated +TIP obtained for 20–24 MT ends per condition. MT end position (0) was determined by staining for β-tubulin. (E and F) Live images of the indicated cell lines transfected with ch-TOG-GFP together with mKate-α-tubulin (larger and smaller fields of view shown; E) and quantification of ch-TOG signal-to-noise ratio at MT tips (F). n = 30 cells in three independent experiments. Mann-Whitney U test.

Article Snippet: We used rabbit antibodies against CAMSAP2 (NBP1-21402, Novus; 17880-1-AP, Proteintech), EB3 , CLIP170 and CLASP1 , GM130 (ab52649; Abcam), GFP (ab290; Abcam), CEP135 (SAB4503685-100UG; Sigma-Aldrich), and goat anti–myosin IIB antibody (sc-47205; Santa Cruz).

Techniques: Disruption, Immunostaining, Control, Expressing, Construct, Incubation, Staining, Transfection, MANN-WHITNEY