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. Author manuscript; available in PMC: 2024 Dec 1.
Published in final edited form as: Biochem Pharmacol. 2023 Oct 29;218:115889. doi: 10.1016/j.bcp.2023.115889

Figure 2: Immunological evidence for the expression of CARD9, BCL-10 and MALT-1 in INS-1 832/13 cells, rat islets and human islets:

Figure 2:

Western blot data depicting expression of CARD9, BCL-10 and MALT-1 in INS-1 832/13 cells, rat islets and human islets is shown here. Methods: INS-1 832/13 cells were propagated in RPMI-1640 medium consisting of 10% FBS supplemented with 100 IU/ ml penicillin and 100 IU/ml streptomycin, 1 mM sodium pyruvate, 50μM 2-mercaptoethanol, and 10 mM HEPES. Prior to a given study, cells were treated overnight with low serum/ low glucose media [79, 80]. Rat (Sprague- Dawley) islets were isolated using the collagenase digestion method as we described in [22, 81]. Human islets (from a 54-year-old Caucasian male, 75”, 172 lbs., with a BMI of 21.6, non-diabetic donor pancreas) were obtained from Prodo Labs (Aliso Viejo, CA, USA). Protocols involving use of rat and human islets received approvals from the appropriate committees at Wayne State University and the JDD VA Medical Center, Detroit. Antibody directed against MALT1 (TA807326S; 1:1,000 dilution) was from OriGene Technologies, Inc. (Rockville, MD, USA). Antibodies against CARD9 (sc-374569; 1:1,000 dilution) and BCL-10 (sc-5273 (1:500 dilution) were from Santa Cruz Biotechnology (Dallas, TX, USA). β-actin antibody (A1978) was from Sigma Aldrich (St. Louis, MO, USA) and used at 1:5,000 dilution. Anti-mouse IgG HRP-conjugated secondary antibody was from Cell Signaling Technology, Inc. (Danvers, MA, USA) used at 1:1,500 dilution. Collectively, these findings suggest expression of CARD9, BCL10 and MALT1 in a variety of insulin secreting cells, including human islets.