Human Reg3A Antibody

Detection of Reg3A by Western Blot

REG3B induces ADM through the RAS-RAF-MEK-ERK signaling pathway.a Western blot showing increased expression of p-ERK, p-MEK, p-BRAF, KRAS, and active RAS in the pancreatic tissue of REG3B-treated WT mice with caerulein-induced pancreatitis (WT cae+REG3B) and a moderate increase in TG mice with caerulein-induced pancreatitis (TG cae) (n = 3, one-way ANOVA). b Western blots demonstrating increased expression of p-ERK, p-MEK, p-BRAF, KRAS, and active RAS in cultured human primary acinar cells and mouse acinar cell line 266-6 after REG3A or REG3B treatment respectively, for 48 h (n = 3, student’s t-test). c Western blot showing the reduced expression of phosphorylated ERK, MEK, BRAF, and total KRAS in the 266-6 cell line after Reg3b gene knockdown by siRNA (n = 3, student t-test). d In the upper panel, LY3009120 inhibited ERK, MEK, BRAF phosphorylation in a dose-dependent manner in 266-6 cell line. In the lower panel, LY3009120 (5 μM) blocked REG3B-induced MEK and ERK phosphorylation (n = 3, one-way ANOVA). e Upper panel, Trametinib efficiently attenuated ERK phosphorylation in a dose-dependent manner in the AR42J cell line (n = 3, one-way ANOVA). Lower panel, Trametinib (100 nM) blocked REG3B-induced ERK phosphorylation. f, g Bright field images (g) with quantification (f) show that Trametinib (100 nM) and LY3009120 (5 μM) treatment hindered REG3B-induced ADM in 3D cultures of mouse primary acinar cells (200×, n = 3, one-way ANOVA). h Confocal microscopy shows that Trametinib (100 nM) and LY3009120 (5 μM) treatment reduced CK19 protein expression (red) and increased AMYLASE protein expression (green) in 3D culture of mouse primary acinar cells (Scale bar: 20 μm). Data are represented as means ± SD, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001. Non-significant (n.s.) if P > 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34099862), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Reg3A by Immunohistochemistry

Increases in oncostatin M (OSM) correlate with elevations of the macrophage chemokines Reg3A and Reg3 gamma in the border zone. Myocardial tissue from six control patients (CON) and six patients with cardiac sarcoidosis (CS) were analyzed by Western blotting (WB) and immunofluorescence (IF). Troponin T (Trop T), actin and hexokinase I (HK I) serve as loading controls. (A) WB and IF images of Reg3A. (B) WB and IF images of Reg3 gamma. (C) IF demonstrates EXTL3 (a potential Reg receptor)-positive granulomata. (D) WB of OSM. (E) IF images of activated cardiomyocytes (P-Stat3, Tyr705). IF images show dedifferentiated non-muscle alpha-actinin 1 (Actn1) expressing cardiomyocytes in the BZ as well as the elongated and sharp shape of cardiomyocytes indicate furthermore OSM activity. The nuclei are stained blue with DAPI. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33923774), licensed under a CC-BY license. Not internally tested by R&D Systems.