Journal: Science Advances

Article Title: Switch of serotonergic descending inhibition into facilitation by a spinal chloride imbalance in neuropathic pain

doi: 10.1126/sciadv.abo0689

Figure Lengend Snippet: ( A ) GFP (green) and PAX2 (red) immunolabeling in the DH. Scale bar, 100 μm. Inset: high magnification showing contacts between Pax2 and 5-HT fibers. Scale bar, 50 μm. ( B ) GFP (green) and Tlx3 (red) immunolabeling in the DH. Scale bar, 100 μm. Inset: high magnification, low or no contact. Scale bar, 25 μm. ( C ) After 3D reconstruction (see Materials and Methods) quantification of contacts between cell bodies (Pax2 and Tlx3) and 5-HT fibers. ( C ) Contacted cells are significantly higher for Pax2(+) than for Tlx3(+) in both males and females [males (blue): 86.2 ± 4.6% for Pax2 and 15.5 ± 3.8% for Tlx3, P < 0.001, Mann-Whitney; females (red): 76.3 ± 2.9% for Pax2 and 8.3 ± 2.2% for Tlx3, P < 0.001, Mann-Whitney]. ( D ) Immunostaining against TPH2 in slices of lumbar spinal cord of GAD67::GFP mice. Scale bar, 25 μm. GABA neurons [GFP(+) in green, stars] or fibers in DH are contacted by TPH2-positive putative synaptic button (in red, white arrows). ( E ) Expression of Myc-tag in synaptic 5-HT terminals in DH following injections of an AAV-synaptophysin-Myc. Immunoreactivity is present in the deeper lamina of the DHSC. Scale bar, 100 μm. Higher magnification shows 5-HT axonic button (white arrow). Scale bar, 20 μm. ( F ) Myc(+) synaptic buttons are in direct contact with GAD67-positive cells and fibers in epet::cre/ GAD67::GFP mice. Scale bar, 20 μm. Inset (top): Myc(+) button on GAD(+) cell bodies. Inset (bottom): Myc(+) button on GAD(+) on GAD(+) fibers.

Article Snippet: For c-Fos experiments, rabbit anti–c-Fos (1:500; 9F6, mAB Cell Signalling Technology 2250 S) was combined with goat anti-Pax2 antibody (1:300; Bio-techne). c-Fos primary antibody was used for 24 hours, and then Alexa Fluor 488–conjugated goat anti-rabbit (1:500; Thermo Fisher Scientific) and Alexa Fluor 647–conjugated donkey anti-goat (1:500; Thermo Fisher Scientific) were used for c-Fos and Pax2, respectively.

Techniques: Immunolabeling, MANN-WHITNEY, Immunostaining, Expressing

Journal: BMC Complementary Medicine and Therapies

Article Title: Toxicity and toxicokinetics of the ethanol extract of Zuojin formula

doi: 10.1186/s12906-022-03684-0

Figure Lengend Snippet: Acquisition parameters in MRM mode

Article Snippet: Carbamazepine (CAS No.: 298–46-4), berberine (CAS No.: 2086-83-1), coptisine (CAS No.: 6020-18-4), epiberberine (CAS No.: 6873-09-2), palmatine (CAS No.: 3486-67-7), jatrorrhizine (CAS No.: 3621-38-3), columbamine (CAS No.: 483–34-1), evodiamine (CAS No.: 518–17-2) and rutaecarpine (CAS No.: 20575–76-2), all > 98% purity, were purchased from Shanghai Yuanye Bio-Technology Company.

Techniques:

Journal: American Journal of Cancer Research

Article Title: MUC1 is responsible for the pro-metastatic potential of calycosin in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Effects of calycosin on the expression of MUC1 (with or without MUC1 gene silencing). Immunofluorescence staining of MUC1 (green) in MIA-PaCa2 cells treated with 100 μM calycosin for 24 h using (A) 2D or (B) 3D cell culture. The nuclei were stained with DAPI (blue). Magnifications: 60× for 2D cell culture (scale bar is 5 μm); 20× for 3D cell culture (scale bar is 50 μm). (C) Gene expression of MUC1 in MIA PaCa-2 cells after treatment with calycosin (50 or 100 μM) for 24 h was detected by quantitative reverse transcription PCR. *P<0.01, **P<0.0001 versus the corresponding negative control (NCtrl) group. #P<0.001, ##P<0.0001 versus corresponding group without calycosin treatment (0 μM). (D) Protein expression of MUC1 in MIA PaCa-2 cells after treatment with calycosin (50 or 100 μM) for 24 h was detected by Western immunoblotting. *P<0.05, **P<0.01, ***P<0.001 versus the corresponding negative control (NCtrl) group. #P<0.05 versus corresponding group without calycosin treatment (0 μM). Cells treated with DMSO were used as negative control. Data are expressed as mean ± SD from 3 independent experiments.

Article Snippet: Calycosin (CAS no. 20575-57-9; >98% purity by HPLC) was purchased from Chengdu Herbpurify Co., Ltd. (Chengdu, Sichuan, China).

Techniques: Expressing, Immunofluorescence, Staining, Cell Culture, Gene Expression, Reverse Transcription, Negative Control, Western Blot

Journal: American Journal of Cancer Research

Article Title: MUC1 is responsible for the pro-metastatic potential of calycosin in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Effects of calycosin on the expression of TGF-β1 (with or without MUC1 gene silencing). Immunofluorescence staining of TGF-β1 (green) in MIA-PaCa2 cells treated with 100 μM calycosin for 24 h using (A) 3D cell culture (scale bar is 50 μm). The nuclei were stained with DAPI (blue). Magnifications: 20× for 3D cell culture. (B) Gene expression of TGF-β1 in MIA PaCa-2 cells after treatment with calycosin (50 or 100 μM) for 24 h was detected by quantitative reverse transcription PCR. #P<0.05 versus corresponding group without calycosin treatment (0 μM). (C) Protein expression of TGF-β1 in MIA PaCa-2 cells after treatment with calycosin (50 or 100 μM) for 24 h was detected by Western immunoblotting. *P<0.05, **P<0.01, ***P<0.001 versus the corresponding negative control (NCtrl) group. (D) The protein interaction of MUC1 and TGF-β in MIA PaCa-2 cells after treatment with calycosin (100 μM) for 24 h was detected by co-immunoprecipitation and Western immunoblotting. *P<0.05, **P<0.01, ***P<0.001 versus the corresponding negative control (NCtrl) group. #P<0.05, ##P<0.01, ###P<0.0001 versus corresponding group without calycosin treatment (0 μM). Cells treated with DMSO were used as negative control. Data are expressed as mean ± SD from 3 independent experiments.

Article Snippet: Calycosin (CAS no. 20575-57-9; >98% purity by HPLC) was purchased from Chengdu Herbpurify Co., Ltd. (Chengdu, Sichuan, China).

Techniques: Expressing, Immunofluorescence, Staining, Cell Culture, Gene Expression, Reverse Transcription, Western Blot, Negative Control, Immunoprecipitation

Journal: American Journal of Cancer Research

Article Title: MUC1 is responsible for the pro-metastatic potential of calycosin in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Effects of calycosin on the migration of MIA PaCa-2 cells (with or without MUC1 gene silencing). A. Wound healing assay was used in MIA-PaCa2 cells treated with 50 μM calycosin with or without MUC1 siRNA for 0, 24, 48 h or 72 h. Area between the inner margins of migrating cells during wound closure was quantified. Three independent experiments were conducted each in triplicate. Data are presented as mean ± SD. *P<0.05, ***P<0.001 versus the corresponding group without calycosin treatment. B. Gene expression of Snail in MIA PaCa-2 cells after treatment with calycosin (50 or 100 μM) for 24 h was detected by quantitative reverse transcription PCR. *P<0.01, **P<0.001 versus the corresponding negative control (NCtrl) group. C. Protein expression of Snail in MIA PaCa-2 cells after treatment with calycosin (50 or 100 μM) for 24 h was detected by Western immunoblotting. *P<0.01, **P<0.001 versus the corresponding negative control (NCtrl) group. #P<0.01 versus corresponding group without calycosin treatment (0 μM). Cells treated with DMSO were used as negative control. Data are expressed as mean ± SD from 3 independent experiments.

Article Snippet: Calycosin (CAS no. 20575-57-9; >98% purity by HPLC) was purchased from Chengdu Herbpurify Co., Ltd. (Chengdu, Sichuan, China).

Techniques: Migration, Wound Healing Assay, Gene Expression, Reverse Transcription, Negative Control, Expressing, Western Blot

Journal: American Journal of Cancer Research

Article Title: MUC1 is responsible for the pro-metastatic potential of calycosin in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Effects of calycosin on the apoptosis of MIA-PaCa2 cells (with or without MUC1 gene silencing). A. FITC/PI staining was performed in MIA-PaCa2 cells treated with 50 μM calycosin with or without MUC1 siRNA for 24 h. Three independent experiments were performed each in triplicate. *P<0.01, **P<0.001 versus the corresponding group without calycosin treatment (No drug). #P<0.01 versus corresponding group without calycosin treatment (0 μM). B. Protein expression of Cleaved-PARP and Atg5 in MIA PaCa-2 cells after treatment with calycosin (50 or 100 μM) for 24 h was detected by Western immunoblotting. *P<0.05, **P<0.01 versus the corresponding negative control (NCtrl) group. #P<0.05 versus corresponding group without calycosin treatment (0 μM). Cells treated with DMSO were used as negative control. Data are expressed as mean ± SD from 3 independent experiments.

Article Snippet: Calycosin (CAS no. 20575-57-9; >98% purity by HPLC) was purchased from Chengdu Herbpurify Co., Ltd. (Chengdu, Sichuan, China).

Techniques: Staining, Expressing, Western Blot, Negative Control

Journal: American Journal of Cancer Research

Article Title: MUC1 is responsible for the pro-metastatic potential of calycosin in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Effects of calycosin on cell cycle modulation in MIA-PaCa2 cells (with or without MUC1 gene silencing). (A) Cell cycle analysis was performed in MIA-PaCa2 cells treated with 100 μM calycosin with or without MUC1 siRNA for 24 h. Gene expression of (B) p27 and (C) p21 in MIA PaCa-2 cells after treatment with calycosin (50 or 100 μM) for 24 h was detected by quantitative reverse transcription PCR. Cells treated with DMSO were used as negative control. Data are expressed as mean ± SD from 3 independent experiments. *P<0.05 versus the corresponding negative control (NCtrl) group. #P<0.01, ##P<0.001, ###P<0.0001 versus corresponding group without calycosin treatment (0 μM).

Article Snippet: Calycosin (CAS no. 20575-57-9; >98% purity by HPLC) was purchased from Chengdu Herbpurify Co., Ltd. (Chengdu, Sichuan, China).

Techniques: Cell Cycle Assay, Gene Expression, Reverse Transcription, Negative Control

Journal: American Journal of Cancer Research

Article Title: MUC1 is responsible for the pro-metastatic potential of calycosin in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Effects of calycosin on metabolic regulation in MIA PaCa2 cells (with or without MUC1 gene silencing). Immunofluorescence staining of AMPKα (red) in MIA-PaCa2 cells treated with 100 μM calycosin for 24 h using (A) 2D or (B) 3D cell culture. The nuclei were stained with DAPI (blue). Magnifications: 60× for 2D cell culture (The scale bar is 5 μm); 20× for 3D cell culture (The scale bar is 50 μm). (C) Gene expression of AMPKα, Sirt1, FGF21 and β-klotho in MIA PaCa-2 cells after treatment with calycosin (50 or 100 μM) for 24 h was detected by quantitative reverse transcription PCR. Cells treated with DMSO were used as negative control. Data are expressed as mean ± SD from 3 independent experiments. *P<0.05, **P<0.01, ***P<0.0001 versus the corresponding negative control (NCtrl) group. #P<0.05, ##P<0.01, ###P<0.0001 versus corresponding group without calycosin treatment (0 μM).

Article Snippet: Calycosin (CAS no. 20575-57-9; >98% purity by HPLC) was purchased from Chengdu Herbpurify Co., Ltd. (Chengdu, Sichuan, China).

Techniques: Immunofluorescence, Staining, Cell Culture, Gene Expression, Reverse Transcription, Negative Control

Journal: American Journal of Cancer Research

Article Title: MUC1 is responsible for the pro-metastatic potential of calycosin in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: The effect of calycosin in PDAC tumor-bearing C57/BL6 mice. A. Calycosin exerted anti-tumor effect in Pan02 tumor-bearing C57/BL6 mice. Ctrl: PBS, PsCtrl: gemcitabine 30 mg/kg i.p. every other day, calcyosin: 30 mg/kg once daily, calycosin + Gun: 100V electroporation for 12 days. More than 80% reduction in tumor mass. B. Daily change of tumor volume over 12 days. Tumor volume of PsCtrl and calycosin groups were decreased, with increase in calycosin + Gun group. C, D. IHC assays of tissues for MUC1, TGF-β, MMP2, MMP9 and CD31 in different treatment groups (magnification =20×).

Article Snippet: Calycosin (CAS no. 20575-57-9; >98% purity by HPLC) was purchased from Chengdu Herbpurify Co., Ltd. (Chengdu, Sichuan, China).

Techniques: Electroporation

Journal: American Journal of Cancer Research

Article Title: MUC1 is responsible for the pro-metastatic potential of calycosin in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: The mechanism of MUC1 in treating PDAC by calycosin.

Article Snippet: Calycosin (CAS no. 20575-57-9; >98% purity by HPLC) was purchased from Chengdu Herbpurify Co., Ltd. (Chengdu, Sichuan, China).

Techniques:

Journal: PeerJ

Article Title: Therapeutic targets and signaling pathways of active components of QiLing decoction against castration-resistant prostate cancer based on network pharmacology

doi: 10.7717/peerj.13481

Figure Lengend Snippet: A list of the active components in Qiling Decoction.

Article Snippet: Formononetin (cat. no. 20100311) was purchased from Jubang Biotechnology Co., Ltd. (Chengdu, China), and Calycosin (20575-57-9) was purchased from Sigma-Aldrich (St. Louis, MO, USA), both of which were dissolved with dimethyl sulfoxide (DMSO).

Techniques:

Journal: PeerJ

Article Title: Therapeutic targets and signaling pathways of active components of QiLing decoction against castration-resistant prostate cancer based on network pharmacology

doi: 10.7717/peerj.13481

Figure Lengend Snippet: In the in vivo experiment, (A) tumor growth was observed, and the (B) volume and (C) weight of tumors were measured; (D) IHC assay was employed to detect the PCNA expression in xenograft tumor. (E) Western blotting was applied to detect the expression levels of HIF-α, VEGFA, TNF- α, IL-6, and P53 in tumor cells treated with QLD and control. (F) PC3 cells were serum-starved, and treated with formononetin for 24 h. Western blotting was employed to detect the expression levels of VEGFA, TNF-α, and IL-6. (G and H) PC3 cells were serum-starved, and treated with formononetin for the indicated time points. (G) CCK-8 and (H) colony formation assays were applied to assess the proliferation of PC3 cells. (I) PC3 cells were serum-starved, and treated with calycosin for 24 h. Western blot and colony formation assays were employed to determine the expression levels of p53, Bax, and C-caspase-3. (J and K) PC3 cells were serum-starved, and treated with calycosin for the indicated time points. (J) CCK-8 and (K) colony formation assays were applied to detect the proliferation of PC3 cells.

Article Snippet: Formononetin (cat. no. 20100311) was purchased from Jubang Biotechnology Co., Ltd. (Chengdu, China), and Calycosin (20575-57-9) was purchased from Sigma-Aldrich (St. Louis, MO, USA), both of which were dissolved with dimethyl sulfoxide (DMSO).

Techniques: In Vivo, Expressing, Western Blot, CCK-8 Assay