pyrrolidinedithiocarbamic acid ammonium pdtc  (Macklin Inc)

Journal: Journal of Inflammation Research

Article Title: Celastrol Mitigates Acute Pancreatitis Associated Inflammation by Modulating the IL-34/CSF-1R Axis and Suppressing NF-κB/ERK Signaling

doi: 10.2147/JIR.S572609

Figure Lengend Snippet: Celastrol attenuates pancreatic injury and inflammatory mediator release in experimental AP. ( A ) Representative H&E staining of pancreatic tissue from Sham, AP, and Celastrol+AP rats with corresponding histological injury scores (scale bar: 100 μm). ( B ) Serum amylase levels (n = 6 rats per group). ( C ) Serum IL-6 and TNF-α levels measured by ELISA (n = 6 rats per group). ( D ) Amylase activity in AR42J cell supernatants under Con (Control), AP (caerulein), and AP plus PD98059, PDTC, or celastrol treatment (n=3 independent experiments). ( E ) IL-6 and TNF-α levels in AR42J cell supernatants (n = 3 independent experiments). Data are presented as mean ± SD. Each dot represents an individual animal or independent experiment. Statistical analysis was performed using one-way ANOVA for data in ( B–E ) with Tukey’s post-hoc test, except for histological scores in ( A ), which were analyzed by Kruskal–Wallis test. *P < 0.05, **P < 0.01, ***P < 0.001 vs AP group.

Article Snippet: To induce acinar-like differentiation, cells were treated with dexamethasone (Solarbio, D8040) for 48 h. After differentiation, cells were stimulated with caerulein (100 nM; MCE, HY-A0190) for 24 h to establish an in vitro AP model. For pharmacological intervention studies, cells were pretreated with celastrol (2.0 μM), PD98059 (10 μM; MCE, HY-12028), or PDTC (60 μM; MCE, HY-18738) for 1 h prior to caerulein stimulation, without removing the inhibitors.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, Control

Journal: Journal of Inflammation Research

Article Title: Celastrol Mitigates Acute Pancreatitis Associated Inflammation by Modulating the IL-34/CSF-1R Axis and Suppressing NF-κB/ERK Signaling

doi: 10.2147/JIR.S572609

Figure Lengend Snippet: IL-34 expression is increased in experimental AP and reduced by celastrol treatment. ( A ) Western blot analysis of IL-34 in pancreatic tissue with densitometric quantification normalized to GAPDH (n = 3 independent biological replicates). ( B ) Serum IL-34 levels measured by ELISA (n = 6 rats per group). ( C ) Representative immunohistochemical staining of IL-34 in pancreatic sections with quantitative analysis (scale bar: 50 μm; n = 6 rats per group). ( D ) Western blot analysis of IL-34 in AR42J cells under Con, AP, and AP plus PD98059, PDTC, or celastrol treatment (n = 3 independent biological replicates). Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 vs AP group.

Article Snippet: To induce acinar-like differentiation, cells were treated with dexamethasone (Solarbio, D8040) for 48 h. After differentiation, cells were stimulated with caerulein (100 nM; MCE, HY-A0190) for 24 h to establish an in vitro AP model. For pharmacological intervention studies, cells were pretreated with celastrol (2.0 μM), PD98059 (10 μM; MCE, HY-12028), or PDTC (60 μM; MCE, HY-18738) for 1 h prior to caerulein stimulation, without removing the inhibitors.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining

Journal: Journal of Inflammation Research

Article Title: Celastrol Mitigates Acute Pancreatitis Associated Inflammation by Modulating the IL-34/CSF-1R Axis and Suppressing NF-κB/ERK Signaling

doi: 10.2147/JIR.S572609

Figure Lengend Snippet: CSF-1R–associated ERK/NF-κB signaling is activated during AP and modulated by celastrol. ( A ) Representative double immunofluorescence staining of CSF-1R (red) and amylase (green) in pancreatic tissue with DAPI nuclear counterstaining (blue). Colocalization indicates spatial association under inflammatory conditions (scale bar: 20 μm). ( B ) Western blot analysis of CSF-1R, p-ERK1/2, and p-p65 in pancreatic tissue (n = 3 independent biological replicates). ( C and D ) Western blot analysis of CSF-1R, p-ERK1/2, and p-p65 in AR42J cells under Con, AP, and AP plus PD98059, PDTC, or celastrol treatment (n = 3 independent biological replicates). The two closely migrating bands detected for p-ERK correspond to the phosphorylated forms of ERK1 (44 kDa) and ERK2 (42 kDa). Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 vs AP group.

Article Snippet: To induce acinar-like differentiation, cells were treated with dexamethasone (Solarbio, D8040) for 48 h. After differentiation, cells were stimulated with caerulein (100 nM; MCE, HY-A0190) for 24 h to establish an in vitro AP model. For pharmacological intervention studies, cells were pretreated with celastrol (2.0 μM), PD98059 (10 μM; MCE, HY-12028), or PDTC (60 μM; MCE, HY-18738) for 1 h prior to caerulein stimulation, without removing the inhibitors.

Techniques: Double Immunofluorescence Staining, Western Blot

Journal: iScience

Article Title: LAR1 promotes breast carcinogenesis by activating NF-κB signaling pathway through binding and enhancing APOC1 expression

doi: 10.1016/j.isci.2025.114386

Figure Lengend Snippet: LAR1 knockdown inhibited p65 nuclear translocation in BC cells (A) The protein expression of p -AKT (Ser473), AKT, p -IKK (Ser180/181), and IKK in MDA-MB-231 cells after LAR1 knockdown and overexpression. (B) The protein expression of p -IκBα (ser32/ser36), IκBα, p-p65 (Ser536), and p65 in MDA-MB-231 cells after LAR1 knockdown and overexpression. (C) Representative images of immunofluorescence staining for p65 in MDA-MB-231 cells after LAR1 knockdown and overexpression. Scale bars, 50 μm. (D) Quantitative analysis for the percentage of nuclear p65-positive cells in images of immunofluorescence staining ( n = 3). (E) Cell viability of MDA-MB-231 cells treated with PDTC (25 μmol/L) for 24 h was detected by CCK-8 assay. (F) Invasion of MDA-MB-231 cells treated with PDTC (25 μmol/L) for 24 h were visualized by Transwell assays ( n = 3). Scale bars, 100 μm. Data were presented as the mean ± SD, with statistical significance assessed using one-way ANOVA (for the left image of D, for E and 7F) or unpaired t test (for the right image of D). ∗∗ p < 0.01, the oeLAR1 group vs. the Vector group. # p < 0.05 and ## p < 0.01, the oeLAR1+PDTC group vs. the oeLAR1+Vehicle group.

Article Snippet: MDA-MB-231 cells were treated with the NF- κB inhibitor PDTC (25 μmol/L, Cat#A800469, Macklin Inc., Shanghai, China) for 24 h. For mRNA stability analysis, MDA-MB-231 cells were treated with 10 μg/mL actinomycin D (Cat#HY-17559, MCE, USA) for different periods of time (0, 1, 2, 3, or 4 h).

Techniques: Knockdown, Translocation Assay, Expressing, Over Expression, Immunofluorescence, Staining, CCK-8 Assay, Plasmid Preparation