Fig 1: circ-CSPP1 knockdown attenuated tumor growth in vivo. (a) Relative circ-CSPP1 expression by qRT-PCR in sh-NC-infected or sh-circ-CSPP1-transduced SW480 cells. (b) Growth curves of the xenograft tumors formed by sh-NC-infected or sh-circ-CSPP1-transduced SW480 cells. (c) Representative images and average weight of the xenograft tumors formed by SW480 cells infected with sh-NC or sh-circ-CSPP1 at day 26 after implantation. qRT-PCR for circ-CSPP1 and miR-431 levels (d), western blot for LASP1 expression (e), and immunohistochemistry assay for Ki-67, MMP9, TUNEL (f) in the xenograft tumors derived from sh-NC-infected or sh-circ-CSPP1-transduced SW480 cells at day 26 after implantation. *P < 0.05, **P < 0.01, ***P < 0.001, or ****P < 0.0001.
Fig 2: LASP1 was a direct target of miR-431 in CRC cells. (a) Schematic of the putative target sequence for miR-431 within LASP1 3′-UTR and mutated miR-431-binding sequence. (b and c) Dual-luciferase reporter assay in both HCT-116 and SW480 cells. (d–f) LASP1 mRNA expression by qRT-PCR and LASP1 protein level by western blot in CRC tissues and matched noncancerous tissues, NCM460, HCT-116, and SW480 cells. (g) Correlation between LASP1 mRNA level and miR-431 expression in CRC tissues using Pearson’s correlation test. Relative miR-431 expression by qRT-PCR (h) and LASP1 protein level by western blot (i) in HCT-116 and SW480 cells transfected with miR-NC mimic, miR-431 mimic, anti-miR-NC, or anti-miR-431. ***P < 0.001 or ****P < 0.0001.
Fig 3: Proteomics analysis of protein expression profiles in rhBMP-2-induced BMSCs. (A) 2D gel electrophoresis maps of BMSCs without or with rhBMP-2 induction (labeled as control and BMP-2, respectively). (B and C) Close-up images of cropped sections of 2-DE maps shown in (A). Protein number 4, LASP1; Number 11 and 12, ferritin light and heavy chains.
Fig 4: Confirmation of RNA and protein expression levels by real-time quantitative PCR and western blotting analysis (n = 3, per treatment type). (A) The expression of LASP1 and ferritin light and heavy chain mRNAs relative to that of GAPDH, the internal control, as detected by real-time PCR. (B) Expression of LASP1 and ferritin light and heavy chains, as assessed by western blotting analysis. The signal corresponding to the protein bands was quantified using densitometric scanning and the relative protein abundance was determined after normalization with the levels of ß-actin protein. Data are presented as mean ± SD and compared using the t-test for independent samples. *p <0.05, indicating significant differences between the two samples compared.
Fig 5: Peptide fingerprint map obtained using MALDI-TOF mass spectrometry. (A) LASP1. (B) Ferritin light chain. (C) Ferritin heavy chain.
Supplier Page from Abcam for Anti-LASP1 antibody