Fig. 5: A 55 kDa peptide from SNRK connects with NOVA1 to affect circ-SNRK formation.
From: Negative feedback of SNRK to circ-SNRK regulates cardiac function post-myocardial infarction
A qRT-PCR analysis of circ-SNRK in NRCMs co-transfected with Ad-SNRK and Ad-NOVA1 or si-SNRK and si-NOVA1. B The expression of NOVA1 mRNA in NRCMs with low- or overexpression of SNRK. C Co-IP of NOVA1 with SNRK in NRCMs. D Co-IP of SNRK with NOVA1 in NRCMs. E Immunoblot analysis of SNRK in NRCMs transfected with pCMV-N-His-SNRK or pCMV-SNRK-C-His using anti-His. F Schematic of SNRK structure. S_TKc Serine/Threonine protein kinases, LCDs low complex domains, D Aspartic acid. G Immunoblot of SNRK and its 55 kDa peptide in NRCMs treated with Z-vad or not. H (Left) Co-IP of activated Caspase 6/7/ 3 with SNRK in NRCMs; (Right) Co-IP of SNRK with activated Caspase 3. I (Left) The level of SNRK and 55 kDa peptide in NRCMs transfected with si-Caspase 3 with time; (Right) Quantification of SNRK level normalized by β-actin via ImageJ. Data were expressed as mean ± SD, n = 3, one-way ANOVA p < 0.05. J The level of circ-SNRK in NRCMs with overexpression of SNRK or 55 kDa peptide. K Confocal analysis of anti-His in NRCMs transfected with pCMV-N-His-SNRK or pCMV-SNRK-C-His. Scale bar = 10 um. L Immunoblot of NOVA1 in Actinomycin D treated NRCMs with low- or overexpression of 55 kDa peptide with time. M RIP of introns flanking circ-SNRK (primer 1, 2, 3) in NRCMs transfected with si-SNRK or pCMV-55kDa peptide. Data were expressed as mean ± SD, n = 3, two-tailed t-test, *p < 0.05 in relative to NC group. A, B, J All data were expressed as mean ± SD, n = 3, two-tailed t-test, *p < 0.05, **p < 0.01, NS not significant.
