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. Author manuscript; available in PMC: 2019 Nov 1.
Published in final edited form as: Ann Rheum Dis. 2018 Jul 30;77(11):1636–1643. doi: 10.1136/annrheumdis-2018-213103

Figure 2. Effect of HK2 modulation in human RA FLS migration and invasion.

Figure 2.

A) RA FLS were transfected with either siRNA control, siRNA HK1 or siRNA HK2 as detailed in methods. Representative images of invasion area (upper panels) and migration (lower panels) are shown. Quantification, as detailed in methods, of area of invasion and migration after PDGF stimulation for 24 hrs are shown. Results are average of 3 different RA FLS lines. Values are the mean ± SEM. B) RA FLS were infected with Ad-GFP, Ad-hHK1 or Ad-hHK2 as detailed in methods. Representative images of invasion area (upper panels) and migration (lower panels) are shown. Quantification, as detailed in methods, of area of invasion and migration after PDGF stimulation for 24 hrs. are shown. Results are average of 3 different RA FLS lines. Values are the mean ± SEM. C) A) qPCR analysis of the indicated genes in RA FLS 72 hrs. after ad-GFP, ad-HK1 and ad-HK2 infection. Results are average of 5 different RA FLS lines. Values are shown as mean ± SEM. * =p<0.05; **=p<0.01. D) left: RNA levels of GLUT1 in RA FLS, 72 hrs. after ad-GFP, ad-HK1 or ad-HK2 infection. Results are average of 3 different RA FLS lines. Values are the mean ± SEM. Right: RA FLS were infected with ad-GFP, ad-HK1 or ad-HK2. Media was changed 48 hrs. after infection and lactate was measured in the supernatants at 2 hrs. and at 4 hrs. Data are pooled from thee technical replicates, and 3 different RA FLS lines. Values are the mean ± SEM. E) RA FLS were infected with Ad-GFP or Ad-hHK2 as detailed in methods. Representing images and quantification are shown of area of invasion after PDGF stimulation for 24 hrs. in the presence or absence of 10uM 7AAC. Results are average of 3 different RA FLS lines. Values are the mean ± SEM. * =p<0.05, **p<0.01, ***p<0.001.