Siscr

We seeded and cultured these cells in six-well plates at 37 °C for 24 h. When the cells reached 50–70% confluence, they were infected with a small interfering EIF3C-1 RNA (si-eIF3c-1, siRNA: 5′-GCAGGACAACATTCAGCAT-3′), a small interfering eIF3c-2 RNA (si-eIF3c-2, siRNA: 5′-GCACACCTACTACAAGTTT-3′) and a negative control siRNA (si-SCR) which were all constructed by RiboBio (Guangzhou, China). Lipofectamine 3000 (Invitrogen) was used for transfections according to the manufacturer’s instructions. The efficiency of the transfections was investigated after 48–72 h using real-time PCR (RT-PCR).

MiR-106b mimics, miR-93 mimics, negative control oligonucleotides (miR-NC), miR-106b inhibitors (anti-miR-106b), miR-93 inhibitors (anti-miR-93), negative control oligo-nucleotide (anti-miR-NC) and PTEN pEGFP-N2 vector (PTEN), empty vector (vector), small interfering RNA of PTEN or Akt (siPTEN, siAkt), scramble siRNA of PTEN or Akt (siSCR) were purchased from RiboBio (Guangzhou, China). MiR-106b and miR-93 level was enhanced by miR-106b mimics or miR-93 mimics and upregulated PTEN was completed by transfection with PTEN in MCF-7 cells. MiR-106b, miR-93 and PTEN were knocked down using anti-miR-106b, anti-miR-93 or siPTEN in MDA-MB-231 cells. Transfection was performed using the Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. The transfection efficiency was evaluated by fluorescence microscopy by calculating the percentage of fluorescein-labeled cells.

For overexpression of HOTAIR and FUT2, HOTAIR or FUT2 cDNA was cloned into the multiple cloning site of the pcDNA3.1 vector (Invitrogen, Carlsbad, CA, USA). MiR-17-5p mimic, negative control oligonucleotides (miR-NC), miR-17-5p inhibitor, negative control oligonucleotide (NC inhibitor), small interfering RNA of HOTAIR or FUT2 (siHOTAIR, siFUT2), scramble siRNA of HOTAIR or FUT2 (siSCR) were purchased from RiboBio (Guangzhou, China). The chondrocytes were seeded into 6-well plates and transfection was performed by using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA). After 48 h of transfection, chondrocytes were stimulated with IL-1β (10 ng/ml) for the 24 h and used for further analysis.

The cDNA of LEF1, circRNF121, and MYD88 was cloned into the multiple cloning site of the pcDNA3.1 vector (Invitrogen, USA). MiR-665 mimic, negative control oligonucleotides (miR-NC), miR-665 inhibitor, negative control oligonucleotide (NC inhibitor), small-interfering RNA of LEF1, circRNF121, or MYD88 (siLEF1, sicirc-1, sicirc-2, sicirc-3, and siMYD88), and scramble siRNA of LEF1 circRNF121 or MYD88 (siSCR) were purchased from RiboBio (Guangzhou, China). Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) was used for the transfection assay in a 6-well plate with seeded chondrocytes. Q-PCR was used to detect the expression of mRNA. Transfection efficiency was detected by fluorescence microscope (OLYPAS, Japan). After 48 h, chondrocytes were stimulated with IL-1β (10 ng/ml) for 24 h and used for further analysis.