Supplementary Table

 S1. AMO AMOs were synthesised with the following modifications: lowercase for DNA, UPPERCASE for 2'OMe, +underlined for LNA, with the asterisks for phosphorothioate linkages. NA (not applicable) denotes AMOs used in Figure 1 -without LNA/DNA counterpart. "z" denotes ZEN groups (see Materials and Methods), "iSp18" refers to an internal spacer, while "3CholMEG" refers to a 3'Cholesterol group. 



  *g*t*+T*c*c*+T*g*c*+T*g*a*+A*c*t*+G*a*g*+C*c*a miR-16-5p CzGCCAAUAUUUACGUGCUGCUz c*g*+C*c*a*+A*t*a*+T*t*t*+A*c*g*+T*g*c*+T*g*c*+T*a miR-34a-5p AzCAACCAGCUAAGACACUGCCz a*+C*a*a*+C*c*a*+G*c*t*+A*a*g*+A*c*a*+C*t*g*+C*c*a miR-29a-3p UzAACCGAUUUCAGAUGGUGCUz t*+A*a*c*+C*g*a*+T*t*t*+C*a*g*+A*t*g*+G*t*g*+C*t*a miR-27a-3p GzCGGAACUUAGCCACUGUGAz g*c*+G*g*a*+A*c*t*+T*a*g*+C*c*a*+C*t*g*+T*g*a*+A miR-21-5p UzCAACAUCAGUCUGAUAAGCUz t*+C*a*a*+C*a*t*+C*a*g*+T*c*t*+G*a*t*+A*a*g*+C*t*a miR-25-3p UzCAGACCGAGACAAGUGCAAUz t*c*+A*g*a*+C*c*g*+A*g*a*+C*a*a*+G*t*g*+C*a*a*+T* 




  




  




  sequences used in this study 


	Human miRNA	AMO sequence 2'OMe	AMO sequence LNA/DNA
	target		
	miR-92a-3p	AzCAGGCCGGGACAAGUGCAAUz	NA
	NC1	GzCGUAUUAUAGCCGAUUAACGzA	NA
	miR-17-5p	CzUACCUGCACUGUAAGCACUUUz	c*+T*a*c*+C*t*g*+C*a*c*+T*g*t*+A*a*g*+C*a*c*+T*t*t*+G
	miR-19a-3p	UzCAGUUUUGCAUAGAUUUGCACz	t*c*+A*g*t*+T*t*t*+G*c*a*+T*a*g*+A*t*t*+T*g*c*+A*c*a
	miR-18a-5p	CzUAUCUGCACUAGAUGCACCUUz	c*+T*a*t*+C*t*g*+C*a*c*+T*a*g*+A*t*g*+C*a*c*+C*t*t*+A
	miR-224-5p	AzACGGAACCACUAGUGACUUz	a*a*+C*g*g*+A*a*c*+C*a*c*+T*a*g*+T*g*a*+C*t*t*+G
	miR-210-3p	UzCAGCCGCUGUCACACGCACAz	t*c*+A*g*c*+C*g*c*+T*g*t*+C*a*c*+A*c*g*+C*a*c*+A*g
	miR-200c-3p	UzCCAUCAUUACCCGGCAGUAUUz	t*+C*c*a*+T*c*a*+T*t*a*+C*c*c*+G*g*c*+A*g*t*+A*t*t*+A
	miR-301a-3p	GzCUUUGACAAUACUAUUGCACUz	g*c*+T*t*t*+G*a*c*+A*a*t*+A*c*t*+A*t*t*+G*c*a*+C*t*g
	miR-331-3p	UzUCUAGGAUAGGCCCAGGGGz	t*+T*c*t*+A*g*g*+A*t*a*+G*g*c*+C*c*a*+G*g*g*+G*c
	miR-125b-5p	UzCACAAGUUAGGGUCUCAGGGz	t*+C*a*c*+A*a*g*+T*t*a*+G*g*g*+T*c*t*+C*a*g*+G*g*a
	miR-146a-5p	AzACCCAUGGAAUUCAGUUCUCz	a*a*+C*c*c*+A*t*g*+G*a*a*+T*t*c*+A*g*t*+T*c*t*+C*a
	miR-126-3p	CzGCAUUAUUACUCACGGUACGz	c*g*+C*a*t*+T*a*t*+T*a*c*+T*c*a*+C*g*g*+T*a*c*+G*a
	miR-134-5p	CzCCCUCUGGUCAACCAGUCACz	c*c*+C*c*t*+C*t*g*+G*t*c*+A*a*c*+C*a*g*+T*c*a*+C*a
	miR-200a-3p	AzCAUCGUUACCAGACAGUGUUz	a*+C*a*t*+C*g*t*+T*a*c*+C*a*g*+A*c*a*+G*t*g*+T*t*a
	miR-191-5p	CzAGCUGCUUUUGGGAUUCCGUUz	c*+A*g*c*+T*g*c*+T*t*t*+T*g*g*+G*a*t*+T*c*c*+G*t*t*+G
	miR-200b-3p	UzCAUCAUUACCAGGCAGUAUUz	t*c*+A*t*c*+A*t*t*+A*c*c*+A*g*g*+C*a*g*+T*a*t*+T*a
	miR-31-5p	AzGCUAUGCCAGCAUCUUGCCz	a*g*+C*t*a*+T*g*c*+C*a*g*+C*a*t*+C*t*t*+G*c*c*+T
	miR-122-5p	CzAAACACCAUUGUCACACUCCz	c*+A*a*a*+C*a*c*+C*a*t*+T*g*t*+C*a*c*+A*c*t*+C*c*a
	miR-100-5p	CzACAAGUUCGGAUCUACGGGUz	c*+A*c*a*+A*g*t*+T*c*g*+G*a*t*+C*t*a*+C*g*g*+G*t*t
	miR-155-5p	AzCCCCUAUCACGAUUAGCAUUAz	a*+C*c*c*+C*t*a*+T*c*a*+C*g*a*+T*t*a*+G*c*a*+T*t*a*a
	miR-182-5p	AzGUGUGAGUUCUACCAUUGCCAAz	a*+G

*t*g*+T*g*a*+G*t*t*+C*t*a*+C*c*a*+T*t*g*+C*c*a*+A*a miR-195-5p GzCCAAUAUUUCUGUGCUGCUz g*+C*c*a*+A*t*a*+T*t*t*+C*t*g*+T*g*c*+T*g*c*+T*a miR-15a-5p CzACAAACCAUUAUGUGCUGCUz c*+A*c*a*+A*a*c*+C*a*t*+T*a*t*+G*t*g*+C*t*g*+C*t*a 
Supplementary Data

Supplementary Figure S1. Dose-dependent inhibition of TLR7/8 by 2'OMe AMOs. Human PBMCs were pre-treated for 45 min with the indicated dose of AMO or RD, and stimulated overnight with 180 nM of B-406AS-1. TNF-α and IFN-α levels were measured in supernatants by ELISA. The data are averaged from three independent experiments in different blood donors, in biological triplicate. Percentages of cytokine production compared to the 0.6 nM RD+B-406AS-1 condition are given. Ordinary one-way ANOVA with Dunnett's multiple comparison tests to the miR-224-5p+B-406AS-1 condition are shown. Unless otherwise indicated, differences were not significant. SEM is shown. S2. Impact of chemistry on 2'OMe AMO inhibitory activity. Human PBMCs (A, B) and mouse primary BMMs (B) pre-treated with 40 nM of the indicated AMO for 45 min were stimulated overnight with 180 nM of B-406AS-1. Human/mouse TNF-α and IFN-α levels were measured in supernatants by ELISA. PS: phosphorothioate modified. CHO: 3'Cholesterol-modified. Percentages of cytokine production compared to the RD+B-406AS-1 condition are given. The data are averaged from a minimum of two independent experiments in three blood donors or three mice, in biological triplicate. Unpaired t-tests comparing miR-224-5p 2'OMe+B-406AS-1 (A) or miR-200a-3p/NC1+B-406AS-1 (B) and indicated conditions are shown ("ns": not significant). 

Supplementary Figure


Supplementary Table S2

MEME analysis of all Class 2 inhibitory 2'OMe AMOs (3'-5' orientation is defined by 'R'), with enriched inhibitory motif and P value (39). S3. FIMO in silico prediction of AMOs with inhibitory motif based on human miRBaseV20 (40). AMO to the indicated miRNAs are in 5'-3' or 3'-5' (indicated with 'R') orientation. 

Supplementary Table