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