Figures & data
Figure 1 kshv-mir-k12-1-5p promotes cell proliferation and inhibits cell apoptosis. (A) The mimics group exhibited increased expression of kshv-mir-k12-1-5p, and the inhibition group exhibited decreased expression of kshv-mir-k12-1-5p. (B and C) Cell proliferation assay was performed by using CCK-8. (D and E) The percentage of apoptosis cells after transfection with kshv-mir-k12-1-5p mimics, mimics NC, inhibitor or inhibitor NC. All data are presented as the mean ± SD. **p<0.01, compared with control groups.
Abbreviation: NC, negative control.
Figure 2 kshv-mir-k12-1-5p promotes cell migration and invasion. (A and B) Cell migration and invasion assays were performed after transfection with kshv-mir-k12-1-5p mimics, mimics NC, inhibitor or inhibitor NC. (C) Micrographs (100×) of 0 and 48 h wound areas were analysed by ImageJ software. (D) Micrographs (200×) of crystal violet-stained cells (invasion cells) were shown . All data are presented as the mean ± SD. **p<0.01, compared with control groups.
Abbreviation: NC, negative control.
Figure 3 kshv-mir-k12-1-5p directly targets the SOCS6 gene. (A) Dual luciferase assay was used to demonstrate the target binding relationship between kshv-mir-k12-1-5p and SOCS6 . (B and C) kshv-mir-k12-1-5p decreased SOCS6 expression at the protein level. (D) kshv-mir-k12-1-5p decreased SOCS6 expression at the mRNA level . All data are presented as the mean ± SD. **p<0.01, compared with control groups.
Abbreviations: SOCS6, suppressor of cytokine signalling 6; NC, negative control; WT, wild type; MUT, mutant type; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.
Figure 4 kshv-mir-k12-1-5p promotes cell growth by targeting SOCS6. (A and B) Western blot was used to detect the protein level of SOCS6 in KS cells transfected with SOCS6 siRNA, kshv-mir-k12-1-5p inhibitor, co-transfected with kshv-mir-k12-1-5p inhibitor and SOCS6 siRNA, respectively. Control: The cells without any transfection. (C) CCK-8 and (D and E) flow cytometry assays were performed to evaluate the proliferation, apoptosis of KS cells. All data are presented as the mean ± SD. **P<0.01 for SOCS6 siRNA vs Control, ^^P<0.01 for kshv-mir-k12-1-5p inhibitor vs co-transfection with kshv-mir-k12-1-5p inhibitor and SOCS6 siRNA.
Abbreviations: siRNA, small interfering RNA; CCK-8, cell counting kit-8.
Figure 5 kshv-mir-k12-1-5p promotes cell metastasis by targeting SOCS6. (A and C) Wound healing and (B and D)Transwell assays were performed to evaluate the migration and invasion of KS cells. All data are presented as the mean ± SD. **P<0.01 for SOCS6 siRNA vs Control, ^^P<0.01 for kshv-mir-k12-1-5p inhibitor vs co-transfection with kshv-mir-k12-1-5p inhibitor and SOCS6 siRNA. (E) Western blot was used to detect the protein level of SOCS6 in KS cells transfected with SOCS6 siRNA, siRNA-NC or co-transfected with kshv-mir-k12-1-5p mimics and SOCS6 siRNA respectively.
Abbreviations: siRNA-NC, small interfering RNA- negative control.
Table S1 The sequences of mimics/inhibitor/NC/siRNA/siRNA NC
Table S2 Primer sequences used for q-PCR
Table S3 kshv-mir-k12-1-5p binding sites in the 3‘ UTR of SOCS6