Figure File Name: 10.1007_s00018-019-03227-w_Fig2

Technique/Assay: Number 9 = adhesion Xcelligence  
MT =Examiner, full name recorded by research group
Instrument Type: Synergy HT microplate reader (BioTek Instruments Inc., VT, USA)
Software version for data generation: Gen5 2.09
Information regarding all further metadata: 

Dataset number 2:

Cells = SUM159, MDA-MB-231
Panel b (bottom)
Replicates: SUM159 1-6, MDA-MB-231 1-4


1: related protocols
2: data analysis Graphpad Prism Version 5.04, serial number: GPW5-101165-NHI-7193 and EXcel 2016, t-test, significance levels *p<0,05, **p<0,01, ***p<0,001, ns: not significant


1) related protocols  

The cell lines SUM159 and MDA-MB-231 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured at 37oC and 5% CO2. 
SUM159 cells were cultured in Hams F12 nutrient mixture (Thermo Fisher Scientific, Waltham, MA USA), supplemented with 5% fetal bovine serum (FBS) and 5 ug/ml bovine pancreas insulin. 
MDA-MB-231 cells were cultured in RPMI 1640 medium. 
In culture media, 10 mM HEPES buffer pH 7.4 and a mixture of 100 U/ml penicillin/100 ug/ml streptomycin (BioConcept Ltd.) was added.
Passage number was between 8-22. 

For transfection of cells with siRNAs, Lipofectamine RNAiMax (Thermo Fisher Scientific) was used. 
Reverse transfection was performed, with the seeding of cells and siRNA transfection taking place simultaneously. 
Per 100,000 cells, 20 pmoles siRNA and 1 ul Lipofectamine reagent was used. 

The target sequences recognized by the siRNAs are: 
mock (scrambled) 5-UGGUUUACAUGUUUUCUGA-3, 
HSD17B12 5-GAACUAAUAUUGUCGGGAA-3 (Microsynth AG, Balgach, Switzerland).

Cells were seeded at a density of 25.000 per well on 96-well plates uncoated or coated with 50 ug/ml fibronectin. 
After incubation for 40 minutes at 37oC, the plate was shaken at 600 rpm for 1.5 minutes. 
Following two washes with PBS, cells were fixed for 12 minutes with 4% formaldehyde. 
Cells were washed twice with PBS and stained with crystal violet for 5 minutes. 1% SDS was then added, and the plate incubated with shaking at 400 rpm for 10 minutes, before absorbance was measured in a spectrum window of 595-600 nm using the Synergy HT microplate reader (BioTek Instruments Inc., VT, USA). 

2) data analysis 

SUM159;;;;;;Normalized to mock w Fibro;;;;
;w Fibro;;wo Fibro;;;w Fibro;;wo Fibro;;
;mock;17B12kd;mock;17B12kd;;mock;17B12kd;mock;17B12kd;
Exp.1;0.79975;0.79325;0.382;0.821;;100;99.18724601;47.76492654;102.6570803;
Exp.2;1.16025;1.14875;1.03625;1.047;;100;99.0088343;89.31264814;90.23917259;
Exp.3;0.353;0.4725;0.382;0.327;;100;133.8526912;;92.63456091;
Exp.4;0.6595;0.735;0.52975;0.625;;100;111.4480667;80.32600455;94.76876422;
Exp.5;0.751666667;0.5875;0.643;0.5285;;100;78.15964523;85.54323725;70.31042129;
Exp.6;0.551666667;0.54;0.533666667;0.40775;;100;97.88519637;96.73716012;73.91238671;
;;;;;;;;;;
;;;;;Avg;;103.2569466;79.93679532;87.42039767;
;;;;;SD;;16.81533623;16.95152907;11.52175936;
;;;;;;;;;;
;;;;;t-test;;0.682987376;;0.476011702;
;;;;;;;;;;
;;;;;;;;;;
MDA-MB-231;;;;;;Normalized to mock w Fibro;;;;
;w Fibro;;wo Fibro;;;;w Fibro;;wo Fibro;
;mock;17B12kd;mock;17B12kd;;;mock;17B12kd;mock;17B12kd
Exp.1;0.81725;0.68325;0.230666667;0.20325;;Exp.1;100;83.60354849;28.22473743;24.86999082
Exp.2;0.737;0.784333333;0.15875;0.16025;;Exp.2;100;106.4224333;21.54002714;21.74355495
Exp.3;0.524333333;0.499333333;0.217;0.2115;;Exp.3;100;95.23204069;41.38588684;40.33693579
Exp.4;0.61475;0.5835;0.1545;0.130666667;;Exp.4;100;94.91663278;25.13216755;21.25525281
;;;;;;;;;;
;;;;;;Avg;;95.04366381;29.07070474;27.05143359
;;;;;;SD;;8.06852304;7.493372926;7.794750614
;;;;;;;;;;
;;;;;;t-test;;0.365389391;;0.757334272