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: After blocking with 5% bovine serum albumin for 1 h at room temperature, sections were incubated overnight at 4 °C with primary antibodies against CSF-1R (1:200) and amylase (Proteintech, 12540-1-AP, 1:200) raised in different host species.
Techniques: Double Immunofluorescence Staining, Western Blot