Journal: Cell and Tissue Research

Article Title: The pro-fibrotic properties of transforming growth factor on human fibroblasts are counteracted by caffeic acid by inhibiting myofibroblast formation and collagen synthesis

doi: 10.1007/s00441-015-2285-6

Figure Lengend Snippet: Characterization of mRNA levels and protein synthesis as observed in non-activated HDFa and HLFa. Fibroblasts were cultured for 48 h in basal medium supplemented with 0.17 mM L-ascorbic acid 2-phosphate magnesium salt and 0.5 % FBS in the absence of TGFβ1 and CAPE. a – g mRNA levels of ACTA2 , TAGLN , GLI1 , GLI2 , COL1A1 , COL1A2 and PLOD2 relative to the reference gene YWHAZ . h – o Representative immunofluorescence stainings ( upper panel ) and p – s quantification of the % of cells ( lower panel ) positive for αSMA, SM22α, collagen type I and LH2. § Statistically significant towards HDFa. Scale bar 100 μm

Article Snippet: HDFa [Caucasian, 20 years, CCD-1093Sk (ATCC ® CRL-2115TM), ATCC, USA] and HLFa [Caucasian, 27 years, CCD-19Lu (ATCC ® CCL-210TM), ATCC, USA] were cultured in basal medium [= Eagle’s minimal essential medium (BE12-662F; Lonza, Switzerland) containing 1 % l-glutamine (Lonza), 1 % penicillin/streptomycin (Gibco Life Technologies, UK)] supplemented with 10 % fetal bovine serum (FBS) (Thermo Scientific, USA).

Techniques: Cell Culture, Immunofluorescence

Journal: Cell and Tissue Research

Article Title: The pro-fibrotic properties of transforming growth factor on human fibroblasts are counteracted by caffeic acid by inhibiting myofibroblast formation and collagen synthesis

doi: 10.1007/s00441-015-2285-6

Figure Lengend Snippet: Effects of TGFβ1 and CAPE on nuclear localization of SMAD2/3. a – c HDFa and d – f HLFa were cultured for 45 min in the presence of TGFβ1 alone, or TGFβ1 in combination with CAPE (co-treatment). Fibroblasts cultured in the presence of TGFβ1 showed a strong nuclear localization of SMAD2/3 ( b , e ), whereas CAPE prevented the TGFβ1-induced nuclear translocation of SMAD2/3 ( c , f ). Scale bar 25 μm

Article Snippet: HDFa [Caucasian, 20 years, CCD-1093Sk (ATCC ® CRL-2115TM), ATCC, USA] and HLFa [Caucasian, 27 years, CCD-19Lu (ATCC ® CCL-210TM), ATCC, USA] were cultured in basal medium [= Eagle’s minimal essential medium (BE12-662F; Lonza, Switzerland) containing 1 % l-glutamine (Lonza), 1 % penicillin/streptomycin (Gibco Life Technologies, UK)] supplemented with 10 % fetal bovine serum (FBS) (Thermo Scientific, USA).

Techniques: Cell Culture, Translocation Assay

Journal: Cell and Tissue Research

Article Title: The pro-fibrotic properties of transforming growth factor on human fibroblasts are counteracted by caffeic acid by inhibiting myofibroblast formation and collagen synthesis

doi: 10.1007/s00441-015-2285-6

Figure Lengend Snippet: Effects of CAPE, TGFβ1 and TGFβ1 + CAPE on ACTA2 mRNA levels and % αSMA-positive cells of HDFa and HLFa. Fibroblasts were cultured for 48 h in the presence of CAPE alone, TGFβ1 alone, or TGFβ1 in combination with CAPE (co-treatment). a , b mRNA levels of ACTA2 relative to the reference gene YWHAZ and expressed as fold-change compared to untreated control (i.e. the baseline level as provided in Fig. ). c – j Representative immunofluorescence stainings ( upper panel ) and k , l quantification of the % of cells ( lower panel ) positive for αSMA. * Statistically significant towards untreated control, and # statistically significant for cells co-treated with TGFβ1 + CAPE towards TGFβ1-treated cells. Scale bar 100 μm

Article Snippet: HDFa [Caucasian, 20 years, CCD-1093Sk (ATCC ® CRL-2115TM), ATCC, USA] and HLFa [Caucasian, 27 years, CCD-19Lu (ATCC ® CCL-210TM), ATCC, USA] were cultured in basal medium [= Eagle’s minimal essential medium (BE12-662F; Lonza, Switzerland) containing 1 % l-glutamine (Lonza), 1 % penicillin/streptomycin (Gibco Life Technologies, UK)] supplemented with 10 % fetal bovine serum (FBS) (Thermo Scientific, USA).

Techniques: Cell Culture, Control, Immunofluorescence

Journal: Cell and Tissue Research

Article Title: The pro-fibrotic properties of transforming growth factor on human fibroblasts are counteracted by caffeic acid by inhibiting myofibroblast formation and collagen synthesis

doi: 10.1007/s00441-015-2285-6

Figure Lengend Snippet: Effects of post-treatment of CAPE on cells treated with TGFβ1 regarding the % αSMA-positive cells of HDFa and HLFa. Fibroblasts were cultured for 48 h in the presence of TGFβ1, followed by a post-treatment with CAPE for 24 h. a – d Representative immunofluorescence stainings ( upper panel ) and e , f quantification of the % of cells ( lower panel ) positive for αSMA. # Statistically significant for cells post-treated with CAPE towards cells stimulated with TGFβ1 only. Scale bar 100 μm

Article Snippet: HDFa [Caucasian, 20 years, CCD-1093Sk (ATCC ® CRL-2115TM), ATCC, USA] and HLFa [Caucasian, 27 years, CCD-19Lu (ATCC ® CCL-210TM), ATCC, USA] were cultured in basal medium [= Eagle’s minimal essential medium (BE12-662F; Lonza, Switzerland) containing 1 % l-glutamine (Lonza), 1 % penicillin/streptomycin (Gibco Life Technologies, UK)] supplemented with 10 % fetal bovine serum (FBS) (Thermo Scientific, USA).

Techniques: Cell Culture, Immunofluorescence

Journal: Cell and Tissue Research

Article Title: The pro-fibrotic properties of transforming growth factor on human fibroblasts are counteracted by caffeic acid by inhibiting myofibroblast formation and collagen synthesis

doi: 10.1007/s00441-015-2285-6

Figure Lengend Snippet: Effects of CAPE, TGFβ1 and TGFβ1 + CAPE on TAGLN mRNA levels and % SM22α-positive cells of HDFa and HLFa. Fibroblasts were cultured for 48 h in the presence of CAPE alone, TGFβ1 alone, or TGFβ1 in combination with CAPE (co-treatment). a , b mRNA levels of TAGLN relative to the reference gene YWHAZ and expressed as fold-change compared to untreated control (i.e. the baseline level as provided in Fig. ). c – j Representative immunofluorescence stainings ( upper panel ) and k , l quantification of the % of cells ( lower panel ) positive for SM22α. * Statistically significant towards untreated control, and # statistically significant for cells co-treated with TGFβ1 + CAPE towards TGFβ1-treated cells. Scale bar 100 μm

Article Snippet: HDFa [Caucasian, 20 years, CCD-1093Sk (ATCC ® CRL-2115TM), ATCC, USA] and HLFa [Caucasian, 27 years, CCD-19Lu (ATCC ® CCL-210TM), ATCC, USA] were cultured in basal medium [= Eagle’s minimal essential medium (BE12-662F; Lonza, Switzerland) containing 1 % l-glutamine (Lonza), 1 % penicillin/streptomycin (Gibco Life Technologies, UK)] supplemented with 10 % fetal bovine serum (FBS) (Thermo Scientific, USA).

Techniques: Cell Culture, Control, Immunofluorescence

Journal: Cell and Tissue Research

Article Title: The pro-fibrotic properties of transforming growth factor on human fibroblasts are counteracted by caffeic acid by inhibiting myofibroblast formation and collagen synthesis

doi: 10.1007/s00441-015-2285-6

Figure Lengend Snippet: Effects of CAPE, TGFβ1 and TGFβ1 + CAPE on COL1A1 and COL1A2 mRNA levels and % collagen type I-positive cells of HDFa and HLFa. Fibroblasts were cultured for 48 h in the presence of CAPE alone, TGFβ1 alone, or TGFβ1 in combination with CAPE (co-treatment). a – d mRNA levels of COL1A1 and COL1A2 relative to the reference gene YWHAZ and expressed as fold-change compared to untreated control (i.e. the baseline level as provided in Fig. ). e – l Representative immunofluorescence stainings ( upper panel ) and m , n quantification of the % of cells ( lower panel ) positive for collagen type I. * Statistically significant towards untreated control, and # statistically significant for cells co-treated with TGFβ1 + CAPE towards TGFβ1-treated cells. Scale bar 100 μm

Article Snippet: HDFa [Caucasian, 20 years, CCD-1093Sk (ATCC ® CRL-2115TM), ATCC, USA] and HLFa [Caucasian, 27 years, CCD-19Lu (ATCC ® CCL-210TM), ATCC, USA] were cultured in basal medium [= Eagle’s minimal essential medium (BE12-662F; Lonza, Switzerland) containing 1 % l-glutamine (Lonza), 1 % penicillin/streptomycin (Gibco Life Technologies, UK)] supplemented with 10 % fetal bovine serum (FBS) (Thermo Scientific, USA).

Techniques: Cell Culture, Control, Immunofluorescence

Journal: Cell and Tissue Research

Article Title: The pro-fibrotic properties of transforming growth factor on human fibroblasts are counteracted by caffeic acid by inhibiting myofibroblast formation and collagen synthesis

doi: 10.1007/s00441-015-2285-6

Figure Lengend Snippet: Effects of post-treatment of CAPE on cells treated with TGFβ1 regarding the % collagen type I-positive cells of HDFa and HLFa. Fibroblasts were cultured for 48 h in the presence of TGFβ1, followed by a post-treatment with CAPE for 24 h. a – d Representative immunofluorescence stainings ( upper panel ) and e , f quantification of the % of cells ( lower panel ) positive for collagen type I. # Statistically significant for cells post-treated with CAPE towards cells stimulated with TGFβ1 only. Scale bar 100 μm

Article Snippet: HDFa [Caucasian, 20 years, CCD-1093Sk (ATCC ® CRL-2115TM), ATCC, USA] and HLFa [Caucasian, 27 years, CCD-19Lu (ATCC ® CCL-210TM), ATCC, USA] were cultured in basal medium [= Eagle’s minimal essential medium (BE12-662F; Lonza, Switzerland) containing 1 % l-glutamine (Lonza), 1 % penicillin/streptomycin (Gibco Life Technologies, UK)] supplemented with 10 % fetal bovine serum (FBS) (Thermo Scientific, USA).

Techniques: Cell Culture, Immunofluorescence

Journal: Cell and Tissue Research

Article Title: The pro-fibrotic properties of transforming growth factor on human fibroblasts are counteracted by caffeic acid by inhibiting myofibroblast formation and collagen synthesis

doi: 10.1007/s00441-015-2285-6

Figure Lengend Snippet: Effects of CAPE, TGFβ1 and TGFβ1 + CAPE on PLOD2 mRNA levels and % LH2-positive cells of HDFa and HLFa. Fibroblasts were cultured for 48 h in the presence of CAPE alone, TGFβ1 alone, or TGFβ1 in combination with CAPE (co-treatment). a , b mRNA levels of PLOD2 relative to the reference gene YWHAZ and expressed as fold-change compared to untreated control (i.e. the baseline level as provided in Fig. ). c – j Representative immunofluorescence stainings ( upper panel ) and k , l quantification of the % of cells ( lower panel ) positive for LH2. * Statistically significant towards untreated control, and # statistically significant for cells co-treated with TGFβ1 + CAPE towards TGFβ1-treated cells. Scale bar 100 μm

Article Snippet: HDFa [Caucasian, 20 years, CCD-1093Sk (ATCC ® CRL-2115TM), ATCC, USA] and HLFa [Caucasian, 27 years, CCD-19Lu (ATCC ® CCL-210TM), ATCC, USA] were cultured in basal medium [= Eagle’s minimal essential medium (BE12-662F; Lonza, Switzerland) containing 1 % l-glutamine (Lonza), 1 % penicillin/streptomycin (Gibco Life Technologies, UK)] supplemented with 10 % fetal bovine serum (FBS) (Thermo Scientific, USA).

Techniques: Cell Culture, Control, Immunofluorescence

Journal: Cell and Tissue Research

Article Title: The pro-fibrotic properties of transforming growth factor on human fibroblasts are counteracted by caffeic acid by inhibiting myofibroblast formation and collagen synthesis

doi: 10.1007/s00441-015-2285-6

Figure Lengend Snippet: Effects of CAPE, TGFβ1 and TGFβ1 + CAPE on GLI1, GLI2 and SNAIL1 mRNA levels of HDFa and HLFa. a – f Fibroblasts were cultured for 48 h in the presence of CAPE alone, TGFβ1 alone, or TGFβ1 in combination with CAPE (co-treatment) on mRNA levels of GLI1, GLI2 and SNAIL1 relative to the reference gene YWHAZ and expressed as fold-change compared to untreated control (i.e. the baseline level as provided in Fig. ). * Statistically significant towards untreated control, # statistically significant for cells co-treated with TGFβ1 + CAPE towards TGFβ1-treated cells

Article Snippet: HDFa [Caucasian, 20 years, CCD-1093Sk (ATCC ® CRL-2115TM), ATCC, USA] and HLFa [Caucasian, 27 years, CCD-19Lu (ATCC ® CCL-210TM), ATCC, USA] were cultured in basal medium [= Eagle’s minimal essential medium (BE12-662F; Lonza, Switzerland) containing 1 % l-glutamine (Lonza), 1 % penicillin/streptomycin (Gibco Life Technologies, UK)] supplemented with 10 % fetal bovine serum (FBS) (Thermo Scientific, USA).

Techniques: Cell Culture, Control