Figure 3. PTRF plays critical role mediating ribosomal transcription response to metabolic challenges in CRISPR/Cas9 genomic editing cell model, without any affect on differentiation.

(A) Indicated protein levels were detected by Western blots from CRISPR/Cas9 genomic edited PTRF null and control 3T3-L1 adipocytes. (B) Relative total TG content and adiponectin secretion levels were measured from CRISPR/Cas9 genomic edited PTRF null and control 3T3-L1 adipocytes. (C) Expression levels changes of indicated proteins during 3T3-L1 differentiation were measured by western blot from CRISPR/Cas9 genomic edited PTRF null and control 3T3-L1 adipocytes. (D) Pre-rRNA levels from nutrients/insulin stimulated (feeding: switching from PBS with 1% BSA to high-glucose DMEM with 10% FBS and insulin) or starved (Fasting: switching the growth medium to PBS with 1% BSA) were measure by RT-qPCR from CRISPR/Cas9 genomic edited PTRF null and control 3T3-L1 adipocytes. (E) After CRISPR/Cas9 genomic edited PTRF null and control 3T3-L1 adipocytes were subject to 'feeding' 12 hr followed by 'fasting' 12 hr cycle for seven days, total TG content, proteins, RNAs and 47S levels were measure as described before. *p<0.05, **p<0.01, and ***p<0.001; Student’s test. Error bars indicate SD.

DOI: http://dx.doi.org/10.7554/eLife.17508.007

Figure 3—figure supplement 1. Characterizations of CRISPR/Cas9 genomic edited PTRF null 3T3-L1 adipocytes.

(A) Whole cell lysates from control and 3 cell lines of CRISPR/Cas9 genomic edited PTRF null 3T3-L1 adipocytes were separated on SDS-PAGE followed by immuno-blots using indicated antibodies. (B) Gene expression levels of some adipocyte specific markers were measured from control and one cell line (KO3) of CRISPR/Cas9 genomic edited PTRF null 3T3-L1 adipocytes by RT-qPCR. (C) Total proteins, RNAs and 47S were measured from control and one cell line (KO3) of CRISPR/Cas9 genomic edited PTRF null 3T3-L1 adipocytes.

DOI: http://dx.doi.org/10.7554/eLife.17508.008