Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 2 Effects of the ginsenoside Rb1 (GRb1) on inflammation and apoptosis in a DSS-induced mouse model of colitis. Expression levels of the colonic cytokines a MPO, b IL-1β, c IL-6, and d TNF-α were determined by ELISA. Western blot analysis of the inflammation-related proteins e iNOS, f COX2, and g p-NF-κB-p65 (calculated as p-NF-κB-p65/NF-κB-p65) and the apoptosis-related protein h cleaved-caspase 3 (cl-caspase 3), calculated as cl-caspase 3/caspase 3. Data are expressed as the mean ± SD. Values in the sham group were set to 100%, and other values are given relative to those in the sham group, **P < 0.01 compared with the sham group; ##P < 0.01 compared with the DSS control group; n = 3 samples for Western blot experiments; n = 6 samples for other experiments.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 3 Effects of the ginsenoside Rb1 (GRb1) on ER stress and Hrd1 in a DSS-induced mouse model of colitis. Western blot analysis of the ER stress marker proteins a GRP78, b PERK, c CHOP, and d caspase 12. e Western blot analysis of Hrd1. f Expression of Hrd1 determined by real- time PCR. g Immunohistochemical staining of Hrd1 in the colonic epithelium. Scale bars, 200, 100, and 50 μm. Data are expressed as the mean ± SD; **P < 0.01 compared with the sham group; ##P < 0.01 compared with the DSS control group; n = 3 samples for Western blot experiments; n = 6 samples for other experiments.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Western Blot, Marker, Expressing, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Staining, Control

Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 5 Effect of the ginsenoside Rb1 (GRb1) on apoptosis. a Effect of GRb1 (200 μM) on IEC-6 cell apoptosis, as determined by flow cytometry. Effect of GRb1 on the expression of b Fas and c caspase 8 in vivo. Effect of GRb1 (200 μM) on the expression of Fas in LPS-treated IEC-6 cells in the absence (d) and presence (e) of siRNA targeting Hrd1. f Effects of GRb1 (200 μM) on IEC-6 cell apoptosis in the presence of siRNA targeting Hrd1. Data are expressed as the mean ± SD. **P < 0.01 compared with the corresponding control group; ##P < 0.01 compared with the DSS control group. n = 3 samples for Western blot experiments, n = 6 samples for other experiments.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Cytometry, Expressing, In Vivo, Control, Western Blot

Journal: Acta pharmacologica Sinica

Article Title: Ginsenoside Rb1 alleviates colitis in mice via activation of endoplasmic reticulum-resident E3 ubiquitin ligase Hrd1 signaling pathway.

doi: 10.1038/s41401-020-00561-9

Figure Lengend Snippet: Fig. 7 Effects of the ginsenoside Rb1 (GRb1) on Hrd1, ER stress, apoptosis, and inflammation in a TNBS-induced colitis model. Western blot analysis of a Hrd1, b CHOP, c GRP78, d cleaved-caspase 3 (cl-caspase 3) (calculated as cl-caspase 3/caspase 3), and e p-NF-κB-p65 (calculated as p-NF-κB-p65/NF-κB-p65). Data are expressed as the mean ± SD. **P < 0.01 compared with the sham group (n = 3 mice); ##P < 0.01 compared with the TNBS-induced colitis group; n = 3 mice.

Article Snippet: Antibodies against Hrd1(13473-1-AP), CHOP(15204-1-AP), Fas (13098-1-AP), caspase 8(13423-1-AP), caspase 3 (19677-1-AP), cleaved-caspase 3 (cl-caspase 3) (19677-1-AP), and iNOS (14142-1-AP) were purchased from Proteintech Co., Ltd. (Wuhan, China).

Techniques: Western Blot

Journal: Journal of Diabetes Investigation

Article Title: Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

doi: 10.1111/jdi.12995

Figure Lengend Snippet: Kbtbd11 expression levels in epididymal white adipose tissue. (a) The expression of Kbtbd11 in epididymal white adipose tissue of C57BL/6J mice; mice were fasted for 24 h or fasted for 24 h/re‐fed for 12 h; n = 3 per group, * P < 0.01 versus fasted. (b) The expression of Kbtbd11 in epididymal white adipose tissue of diet‐induced obesity mice; diet‐induced obesity mice were fed a high‐fat diet for 1 month. The mice were fasted for 24 h; n = 3 per group, * P < 0.01 versus chow.

Article Snippet: Furthermore, Kbtbd11 knockdown inhibited adipogenesis, but only before MCE (not mature 3T3‐L1 adipocytes), and Kbtbd11 overexpression induced MCE, leading to the expression of Cebpa and Pparg (Figure c,f).

Techniques: Expressing

Journal: Journal of Diabetes Investigation

Article Title: Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

doi: 10.1111/jdi.12995

Figure Lengend Snippet: Kbtbd11 messenger ribonucleic acid (mRNA) expression in differentiating 3T3‐L1 cells. Total RNA was extracted from 3T3‐L1 cells at day 0, 2, 4, 8 and 10 after the induction of differentiation by treatment with an adipogenic cocktail; n = 3 per group, * P < 0.01 versus day 0.

Article Snippet: Furthermore, Kbtbd11 knockdown inhibited adipogenesis, but only before MCE (not mature 3T3‐L1 adipocytes), and Kbtbd11 overexpression induced MCE, leading to the expression of Cebpa and Pparg (Figure c,f).

Techniques: Expressing

Journal: Journal of Diabetes Investigation

Article Title: Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

doi: 10.1111/jdi.12995

Figure Lengend Snippet: Kbtbd11 messenger ribonucleic acid (mRNA) expression in response to adipogenic stimuli at early time points. 3T3‐L1 preadipocytes were cultured to confluence, serum‐starved for 12 h and treated with (a) 5 μg/mL insulin, (b) 2.5 μmol/L dexamethasone (DEX) or (c) 200 μmol/L 3‐isobutyl‐1‐methylxanthine (IBMX) for the indicated times; n = 3 per group, * P < 0.01 versus 0 h.

Article Snippet: Furthermore, Kbtbd11 knockdown inhibited adipogenesis, but only before MCE (not mature 3T3‐L1 adipocytes), and Kbtbd11 overexpression induced MCE, leading to the expression of Cebpa and Pparg (Figure c,f).

Techniques: Expressing, Cell Culture

Journal: Journal of Diabetes Investigation

Article Title: Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

doi: 10.1111/jdi.12995

Figure Lengend Snippet: The effects of knockdown and overexpression of Kbtbd11 on 3T3‐L1 cellular differentiation. (a) The expression of Kbtbd11 messenger ribonucleic acid (mRNA) in Kbtbd11 knockdown 3T3‐L1 cells at day 8. Kbtbd11 knockdown adenovirus particles were used with either of the two independent Kbtbd11 short hairpin ribonucleic acid (shRNA) constructs (shRNA#1 and shRNA#2); n = 3 per group, * P < 0.01 versus LacZ‐specific short hairpin ribonucleic acid (shLacZ); (b) triglyceride accumulation in 3T3‐L1 cells on day 8, visualized using Oil Red O staining; (c) the mRNA levels in 3T3‐L1 cells expressing each shRNA at various time points after inducing differentiation; n = 3 per group, * P < 0.01 versus shLacZ; (d) the expression of Kbtbd11 mRNA in Kbtbd11 ‐overexpressing 3T3‐L1 cells at day 8. Cells were infected with adenoviral vectors for expressing green fluorescent protein (GFP) or mouse Kbtbd11 ; n = 3 per group, * P < 0.01 versus GFP; (e) triglyceride accumulation in 3T3‐L1 cells on day 6 was detected using Oil Red O staining. (f) Relative mRNA levels in each group of 3T3‐L1 cells at various time points after inducing differentiation; n = 3 per group, * P < 0.01 versus GFP.

Article Snippet: Furthermore, Kbtbd11 knockdown inhibited adipogenesis, but only before MCE (not mature 3T3‐L1 adipocytes), and Kbtbd11 overexpression induced MCE, leading to the expression of Cebpa and Pparg (Figure c,f).

Techniques: Knockdown, Over Expression, Cell Differentiation, Expressing, shRNA, Construct, Staining, Infection

Journal: Journal of Diabetes Investigation

Article Title: Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

doi: 10.1111/jdi.12995

Figure Lengend Snippet: Effects of knockdown of Kbtbd11 on mature 3T3‐L1 adipocytes. (a) Triglyceride accumulation in Kbtbd11 ‐knockdown mature 3T3‐L1 adipocytes at 48 h after either of the two independent Kbtbd11 short hairpin ribonucleic acid (shRNA) adenoviral infections (shRNA#1 and shRNA#2) visualized using Oil Red O staining; (b) RNA was harvested at 48 h after adenoviral infection, and expression levels of Kbtbd11 , adipocytes ( Pparg and aP2 ) and inflammation markers ( Tnfa and Il6 ), and lipogenic ( Fasn ) and proapoptotic genes ( Bax and Bcl2 ) were measured using quantitative polymerase chain reaction; n = 3 per group, * P < 0.01 versus shLacZ.

Article Snippet: Furthermore, Kbtbd11 knockdown inhibited adipogenesis, but only before MCE (not mature 3T3‐L1 adipocytes), and Kbtbd11 overexpression induced MCE, leading to the expression of Cebpa and Pparg (Figure c,f).

Techniques: Knockdown, shRNA, Staining, Infection, Expressing, Real-time Polymerase Chain Reaction