hACVR1C(3)

Fig.1 Detection of ACVR1C expression in adipose tissues by RT-PCR. Total RNA of eWAT, mammary gland and BAT was extracted from WT C57BL/6 mice and homozygous hACVR1C(3) knockin mice (n=3, male, 6 weeks old), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human mRNA primers.

Abbr. M, marker; HO, homozygous; WT, wild type; eWAT, epididymal adipose tissue; BAT, brown adipose tissue.

Fig.2 Detection of human ACVR1C (A) and mouse Acvr1c (B) expression in adipose tissues by qPCR. Total RNA of eWAT, mammary gland and BAT was extracted from WT C57BL/6 mice and homozygous hACVR1C(3) knockin mice (n=3, male, 6 weeks old), then cDNA libraries were synthesized by reverse transcription, followed by qPCR with human ACVR1C-, mouse Acvr1c- and mouse Gapdh-specific primers. Relative expression represents human ACVR1C (A) or mouse Acvr1c (B) mRNA level relative to its average expression in hACVR1C(3) knockin mice or WT mice, respectively. 

Abbr. HO, homozygous; WT, wild type; eWAT, epididymal adipose tissue; BAT, brown adipose tissue.

Fig.3 Detection of ACVR1C expression in hACVR1C(3) knockin mice by WB. Tissue lysates of mammary gland and brown adipose tissue (BAT) were collected from wild-type C57BL/6 mice (WT) and homozygous hACVR1C(3) knockin mice (HO) (male, 6 weeks old), and then analyzed by western blot with anti-ACVR1C antibody (Santa Cruz, sc-374538). 20 μg of total proteins were loaded for western blotting analysis. The antibody is cross-reactive between human and mouse ACVR1C.

Fig.4 Detection of human ACVR1C mRNA knockdown in adipocyte tissues by qPCR. 10-week-old, female and male homozygous hACVR1C(3) mice were assigned to two groups: a vehicle control group (n=4, 3 female and 1 male) and a test drug group (n=4, 2 female and 2 male). Following a single subcutaneous dose of vehicle or a small nucleic acid drug from a client, mice were sacrificed at day 14 for tissue collection. Expression levels of human ACVR1C mRNA in iWAT, eWAT, and BAT were determined via qPCR, with data normalized against the endogenous mouse Ppia reference gene (Mean±SEM, T-test, ns, no significance). (In cooperation with a client).