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. Author manuscript; available in PMC: 2022 Apr 13.
Published in final edited form as: FASEB J. 2022 Mar;36(3):e22192. doi: 10.1096/fj.202101298R

FIGURE 5.

FIGURE 5

Jmjd3 promotes muscle differentiation in vitro. (A) QPCR data showing relative expression of Jmjd3 in sub-confluent (70%) and confluent (100%) myoblasts and in myogenic cells at 1 day, 5 days, and 7 days following the onset of differentiation. *Indicates significantly different from 70% confluent myoblast control group at p < .05 analyzed by one-way ANOVA with Dunnett’s multiple comparison test. Error bar represents SEM. N = 5 for each data set. (B) Western blot showing relative expression of Jmjd3 and KL in sub-confluent myoblasts and differentiated myotubes. (C, D) QPCR data showing relative expression of Myod1 (C) and Myog (D) in myogenic cell cultures. *Indicates significantly different from 70% confluent myoblast control group at p < .05 analyzed by one-way ANOVA with Dunnett’s multiple comparison test. Error bar represents SEM. N = 5 for each data set. (E–H) QPCR data showing relative expression for Jmjd3 (E), Pax7 (F), Myod1 (G), and Myog (H) in cultured myoblast cells transfected with control siRNA or siRNA targeting Jmjd3. *Indicates significantly different from cells transfected with control siRNA at p < .05 analyzed by t-test. Error bar represents SEM. N = 3 for all data sets. (I–L) QPCR data showing relative expression for transcripts of Jmjd3 (I), Pax7 (J), Myod1 (K), and Myog (L) in cultured myoblasts transfected with control pCS2-vector plasmid or pCS2-Jmjd3-F expression plasmid for 6-h followed by 72-h in differentiation conditions. *Indicates significantly different from cells transfected with control pCS2-vector plasmid at p < .05 analyzed by t-test. Error bar represents SEM. N = 3 for all data sets. (M) Western blot showing relative levels of Jmjd3 and myogenin in myogenic cells transfected with control pCS2-vector plasmid or pCS2-Jmjd3-F expression plasmid for 6-h followed by 72-h in differentiation conditions. (N, O) Quantification of total Jmjd3 protein (N) or myogenin protein (O) relative to protein loaded per gel lane. *Indicates significantly different from cells transfected with control pCS2-vector plasmid at p < .05 analyzed by t-test. Error bar represents SEM. N = 3 for all data sets