Fig 2: miR-877 inhibits CRC tumor growth in vivo. (A) Tumor xenografts derived from SW480 cells transfected with miR-877 mimics or miR-NC. (B) Tumor length and width were measured every 4 days, and tumor volumes were calculated using the following formula: tumor volume (mm3) = width2 (mm2) × length (mm)/2. *P<0.05 versus miR-NC. (C) After 30 days, all mice were sacrificed, and xenografts were excised and weighed. *P<0.05 versus miR-NC. (D) RT-qPCR was utilized to determine miR-877 expression in tumor xenografts derived from SW480 cells transfected with miR-877 mimics or miR-NC. *P<0.05 versus miR-NC. (E) Expression levels of MTDH, PTEN, p-Akt, and Akt in tumor xenografts were detected using western blotting analysis.

Fig 3: Effects of bta-miR-199a-3p on the LPS-induced PI3K/AKT/NF-?B pathway. (A,B) Effects of overexpression or inhibition of bta-miR-199a-3p on the mRNA expression levels of TLR4, NFKB1, and RELA. (C,D) Effects of overexpression or inhibition of bta-miR-199a-3p on the mRNA expression levels of PIK3R1, PIK3CA, AKT1, and AKT3. (E–G) Effects of overexpression or inhibition of bta-miR-199a-3p on the protein expression levels of PI3K, AKT, p65, and p-p65: (E) is the result of Western blot, and (F,G) are the image analysis results of (E) (with ß-actin as a standardized correction). * refers to p < 0.05, ** refers to p < 0.01, ns refers to p > 0.05.

Fig 4: Western blotting analysis for Akt, FoxO1, Pdx1, MafA, and MafB in rat islets. GAPDH was used as a loading control (#P < 0.05vs. NC; *P < 0.05vs. DM; •P < 0.05vs. GLP1 50 µg/kg; and ?P < 0.05vs. GLP1 100 µg/kg). GLP1, glucagon-like peptide 1; NC, negative control; DM, diabetes mellitus; Akt, AKT kinase; FoxO1, forkhead box O 1; Pdx1, pancreatic and duodenal homeobox 1; MafA, MAF BZIP transcription factor A; MafB, MAF BZIP transcription factor B; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Fig 5: Signalling pathways leading to apoptosis. Western Blot data based on literature research36,37 after transfection of SKOV3 cells with miRNA-744 or with siRNA against NFIX (a) or HNRNPC (b) and qRT-PCR (c) confirmed the potential signalling pathway by reduced expression of NFIX and Bcl2 as well as diminished phosphorylation of AKT. By binding to NFIX and HNRNPC miRNA-744 is interfering with the intrinsic apoptotic pathway. Downregulation of HNRNPC resulted in enhanced expression and activation of PDCD4 and p70S6K36 (e). Reduced NFIX expression, induced by binding of miR-744, leads to reduced expression of Bcl2 leading to apoptosis (d). The expression analysis of miR-744 and miR-21 was normalized to the CT value of U6 snRNA55. Statistical differences were tested using paired t-test. In order to analyse DNA fragmentation (cells in sub G0/G1, d) SKOV3 cells were seeded and transfected as described in Fig. 3 with miR-744 mimics, non-targeting siRNA (NT, negative control for cell death), or siRNA against NFIX or HNRNPC. Statistical analysis was performed by one-way ANOVA followed by Bonferroni post-test (d). [n = 3 technical replicates; mean ± SD; **p < 0.01; ***p < 0.001; ****p < 0.0001]. The cells were harvested for Western Blot analysis 60 h after treatment (a and b). GAPDH was used as loading control.