Bfc regulates the expression of Crq in vivo.

A. The relative crq mRNA levels in stage 7–14 wild type embryos were quantified by qPCR; significant differences were observed compared to the expression in stage 7 embryos. B. The bfc mRNA levels in stage 7–14 wild-type embryos were quantified by qPCR; significant differences were observed compared to the expression in stage 7 embryos. C. The relative crq mRNA levels were quantified and compared in the same stage of wild type and bfc^ko mutant embryos by qPCR. D. Western blot analysis was used for the detection of the Crq protein levels in stage 11–14 embryos of wild type and bfc mutant flies. Actin was used as the loading control. E. The relative Crq protein levels in stage 11–14 wild type, and bfc^ko mutant embryos were analyzed via western blotting. The protein levels were normalized to those of actin. Significant differences were observed in comparison with stage 11 wild-type embryos; n = 3. F. The relative crq and bfc mRNA levels were quantified by qPCR in stage to 10–13 wild-type w¹¹¹⁸ and H99 (absence of apoptosis) embryos. G. The crq, bfc, and drpr mRNA levels were quantified by qPCR and normalized to the Rpl32 mRNA levels. “*”/“#”/“※” represent the significant differences in the context of crq/bfc/drpr expression, respectively. “##” indicates P<0.01; “###” indicates P<0.001; “####” indicates P<0.0001; “※※” indicates P<0.01. H. The drpr mRNA levels were quantified by qPCR in wild-type S2 cells and bfc-RNAi S2 cells in the presence or absence of ACs. Statistical significance was assessed using the one-way ANOVA(A, B, E, F, H) or two-way ANOVA (C, G).

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