human rh tgf β1  (R&D Systems)

Journal: Arthritis Research & Therapy

Article Title: TGFβ signaling pathway is altered by HLA-B27 expression, resulting in pathogenic consequences relevant for spondyloarthritis

doi: 10.1186/s13075-024-03370-1

Figure Lengend Snippet: BMP/TGFβ pathway receptors expression in Tn and Teff from B27 and NTG rats. The mRNA expression of TGFβ and activin receptor genes was assessed by RT-qPCR in ex-vivo sorted Tn and Teff from NTG (grey bar – white dots) and B27 (red bar – red dots) adult rat mLNs. Scattered dot plot with bars represents the mean of mRNA relative expression in arbitrary units (A.U) out of 8 to 11 experiments. Differences between NTG and B27 expression levels for each receptor were tested by unpaired t-tests with Welch’s correction (**: p < 0.01). Error bars show SEM. On the first row Acvr1 , Bmpr1a , Acvr1b , Tgfbr1 and Acvr1c genes coding for ALK2, ALK3, ALK4, ALK5 and ALK7 BMPR1s respectively. On the second row Acvr2a , Acvr2b , Bmpr2 and Tgfbr2 represent type 2 BMP receptor genes

Article Snippet: Single-cell suspensions (10 6 cells) from mLN were stimulated with recombinant human (rh) TGFβ1 (5ng/ml) (Miltenyi Biotec) or PBS for 1 h at 37 °C/5% CO2.

Techniques: Expressing, Quantitative RT-PCR, Ex Vivo

Journal: Arthritis Research & Therapy

Article Title: TGFβ signaling pathway is altered by HLA-B27 expression, resulting in pathogenic consequences relevant for spondyloarthritis

doi: 10.1186/s13075-024-03370-1

Figure Lengend Snippet: Basal pSMAD2/3 is reduced in B27 rat T cells but increases more after TGFβ1 stimuation. ( A ) Left: Representative histograms of pSMAD2/3 expression in mLN T cells from NTG and B27 rats after 1 h exposure to medium alone (PBS) or TGFβ1 at 37 °C. Right: Intracellular pSMAD2/3 was quantified in mLN T cells from B27 and NTG adult rats after exposure to TGFβ1 or medium alone. Scatter dot plots with bars representing the mean of the SI of 8 experiments. Similar symbols indicate paired conditions. Vertical error bars show the SEM. Paired t-test *: p < 0.05. ( B , C ) Intracellular pSMAD2/3 was quantified in Tn ( B , C ) or sorted Teff ( C ) from B27 and NTG adult rats after exposure to medium (PBS) or TGFβ1 in 8 to 9 independent experiments. Similar symbols indicate paired conditions. ( B ) Bars height shows the SI mean values. Vertical error bars show the SEM. Paired t-test *: p < 0.05. ( C ) Mean of pSMAD2/3-fold change after exposure to TGFβ expressed as the ratio of pSMAD2/3 SI after TGFβ1 stimulation over the SI without exposure from ( B ) (left panel) and from sorted Teff (right panel). Paired t-test *: p < 0.05; ***: p < 0.001, ( D ) Intracellular pSMAD2/3 was quantified after TGFβ1 stimulation or no stimulation by gating on Tn or Teff from premorbid NTG and B27 rats. Scatter dot plots with bar show the pSMAD2/3 SI. Similar symbols indicate paired conditions. Bars height show the SI mean values ( n = 7–9 independent experiments). Vertical error bars show the SEM. Paired t-test *: p < 0.05

Article Snippet: Single-cell suspensions (10 6 cells) from mLN were stimulated with recombinant human (rh) TGFβ1 (5ng/ml) (Miltenyi Biotec) or PBS for 1 h at 37 °C/5% CO2.

Techniques: Expressing

Journal: Arthritis Research & Therapy

Article Title: TGFβ signaling pathway is altered by HLA-B27 expression, resulting in pathogenic consequences relevant for spondyloarthritis

doi: 10.1186/s13075-024-03370-1

Figure Lengend Snippet: TGFβ target genes are differentially expressed between NTG and B27 rat Tn. ( A ) mRNA relative expression in mLN-sorted Tn from NTG (grey bar – white dots) or B27 (red bar – red dots) adult rats immediately after sorting (T0) or after 1 h, 3 h and 6 h of incubation, without (PBS) or with TGFβ1, assessed by RT-qPCR. Scattered dot plots with bars show the mean of the mRNA relative expression out of 6 NTG and 8 B27 experiments. ( B ) Tgfb1 mRNA relative expression in ex-vivo sorted Tn from NTG and B27 adult (left panel) and premorbid (right panel) rats was assessed by RT-qPCR. Scattered dot plot with bars show the mean of mRNA relative expression out of 10 experiments for adult rats and 8 experiments for premorbid rats. In all graphics, differences between NTG and B27 expression level was tested by Welch’s t-tests, *: p < 0.05; **: p < 0.01; ***: p < 0.001

Article Snippet: Single-cell suspensions (10 6 cells) from mLN were stimulated with recombinant human (rh) TGFβ1 (5ng/ml) (Miltenyi Biotec) or PBS for 1 h at 37 °C/5% CO2.

Techniques: Expressing, Incubation, Quantitative RT-PCR, Ex Vivo