Fig 1: TNF-α upregulated RGS16 expression in PBMCs. PBMCs were stimulated with TNF-α (20 ng/mL), IL-4 (20 ng/mL), IL-6 (20 ng/mL), IL-12 (20 ng/mL), IL-23 (20 ng/mL) and lipopolysaccharide (LPS) (200 ng/mL) for 8 h, respectively. Expression of RGS16 mRNA was detected by real-time reverse transcription-PCR (qRT-PCR). *P < 0.05, **P < 0.01, ***P < 0.001 versus control. (b) PBMCs were stimulated with different concentrations of TNF-α for 8 h, and expression of RGS16 mRNA was detected by qRT-PCR. *P < 0.05, **P < 0.01, ***P < 0.001 versus control. (c) PBMCs were stimulated with TNF-α (20 ng/mL) for 4, 8 and 12 h, and expression of RGS16 mRNA was detected by qRT-PCR. ***P < 0.001 versus control. (d) PBMCs were pretreated with caspase-3/8 inhibitor (Z-DEVD-FMK, 50 μM), JNK inhibitor (JNK-IN-7, 1 μM), or NF-κB inhibitor (BAY 11-7082, 1 μM) followed by TNF-α (20 ng/ml), respectively. Expression of RGS16 mRNA was detected by qRT-PCR. **P < 0.01, ***P < 0.001 versus control. PBMCs, peripheral blood mononuclear cells; RGS, regulator of the G-protein signaling.
Fig 2: RGS16 expression significantly increased in PBMCs from patients with ulcerative colitis (UC). (a) RGS16 mRNA expression in PBMCs from patients with UC (n = 22), and healthy controls (HC) (n = 15) was examined by real-time reverse transcription-PCR (qRT-PCR). Gene expression was normalized to GAPDH. **P < 0.01. (b–d) Correlation analysis was performed between ESR (b), serum TNF-α (c) or IL-17A levels (d) and RGS16 mRNA expression in PBMCs from patients with UC. ESR, erythrocyte sedimentation rate; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PBMCs, peripheral blood mononuclear cells; RGS, regulator of the G-protein signaling.
Fig 3: Anti-TNF treatment downregulated RGS16 expression. (a) Freshly obtained inflamed colonic tissue from patients with active ulcerative colitis (UC) (n = 12) were treated with IFX (50 μg/mL) or control for 24 h, and RGS16 mRNA expression was measured by qRT-PCR. (b) PBMCs isolated from UC patients (n = 16) were stimulated with IFX (50 ng/ml) for 24 h, and RGS16 mRNA expression was measured by qRT-PCR. *P < 0.05, **P < 0.01. IFX, infliximab; PBMCs, peripheral blood mononuclear cells; RGS, regulator of the G-protein signaling.
Fig 4: RGS16 expression increased significantly in inflamed mucosa from patients with ulcerative colitis (UC). (a) Representative images of immune-histochemical staining of RGS16 expression in inflamed colon from patients with UC and normal colonic mucosa from healthy controls (HC). Scar bar represents 50 μm. (b) Percentages of RGS16+ cells in colonic mucosa in a were shown in the bar. **P < 0.01. (c) Colon biopsy specimens were collected from patients with UC (n = 28), and HC (n = 22). RGS16 mRNA expression was detected by using real-time reverse transcription-PCR (qRT-PCR). Gene expression was normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in each group. ***P < 0.001. (d) RGS16 protein expression in colonic mucosa from patients with UC (n = 11), and HC (n = 9) was examined by western blot, with GAPDH as reference. (e) Relative expression of RGS16 protein expression in D. **P < 0.01. (f) RGS16 mRNA expression in inflamed and unaffected intestinal mucosa from the same patients with UC (n = 12) was examined by qRT-PCR. ***P < 0.001. (g). RGS16 protein expression in inflamed and unaffected intestinal mucosa from the same patients with UC (n = 8) was examined by western blot. (h) Relative expression of RGS16 protein expression in g. **P < 0.01. RGS, regulator of the G-protein signaling.
Fig 5: Increased RGS16 expression in colonic mucosa was correlated with disease activity. (a) Correlation analysis was performed between Mayo index and RGS16 mRNA expression in inflamed mucosa of patients with ulcerative colitis (UC) (***P < 0.001). (b) RGS16 mRNA expression in colonic mucosa from patients with slight UC (n = 9), mild UC (n = 13), severe UC (n = 6) and healthy controls (HC) (n = 15) was analyzed. Gene expression was normalized to GAPDH in each group. *P < 0.05, **P < 0.01, ***P < 0.001. (c) Correlation analysis was performed between UCEIS and RGS16 mRNA expression in inflamed mucosa of patients with UC.***P < 0.001. (d,e) RGS16 expression was examined in colonic mucosa from patients after or before the treatment of 5-ASA (d, n = 15) or prednisone (e, n = 9). 5-ASA, 5-aminosalicylic acid; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; RGS, regulator of the G-protein signaling; UCEIS, ulcerative colitis endoscopic index of severity.
Supplier Page from Abcam for Anti-RGS16 antibody [OTI4E5]