Figure 1.
Figure 1. Expression of major ECC transcripts and proteins in human orbicularis oculi muscle biopsies. (A) Gene expression was performed by qPCR as described in Materials and methods. Each reaction was performed in triplicate, in pooled muscle samples from five biopsies from different individuals. Expression levels were normalized to ACTN2 expression. Results are expressed as mean fold change of transcripts in orbicularis oculi compared with quadriceps; the latter was set as 1. Results were analyzed using the Student’s t test: *, P < 0.05; ***, P < 0.0001. (B) MYH13 gene expression was performed by qPCR as described in Materials and methods. Each reaction was performed in triplicate, in pooled muscle samples from five biopsies from different individuals. Expression levels were normalized to ACTN2 expression. Results are expressed as mean fold change of transcripts in orbicularis oculi (OO) compared with quadriceps (QU); the latter were set as 1. Results were analyzed using the Student’s t test: ***, P < 0.0001. The inset shows MyHC protein expression in EOM and orbicularis oculi muscle homogenates; 10 and 30 µg protein were loaded per lane, respectively. Gels and conditions were as described in Materials and methods. (C) Western blot analysis of total SR proteins in human orbicularis oculi and quadriceps muscles. 30 µg of protein was loaded per lane and separated on 6% or 10% SDS-PAGE. Blots were probed with the indicated antibodies. Bar histograms represent the mean (±SEM) band intensity normalized to SRL content. *, P < 0.05. (D) Western blot of human quadriceps, EOM, orbicularis oculi, and mouse heart (positive control) total SR proteins, probed with Cav1.2 antibody. 30 µg of protein was loaded per lane and separated on 6% SDS-PAGE. (E) Western blot of total muscle extracts; 30 µg of protein was loaded per lane, separated on 6% SDS-PAGE, and probed with anti-utrophin (top) and anti-dystrophin (bottom) antibodies. Bar histograms represent the mean (±SEM) band intensity normalized to MyHC content. Student’s t test: *, P < 0.05.

Expression of major ECC transcripts and proteins in human orbicularis oculi muscle biopsies. (A) Gene expression was performed by qPCR as described in Materials and methods. Each reaction was performed in triplicate, in pooled muscle samples from five biopsies from different individuals. Expression levels were normalized to ACTN2 expression. Results are expressed as mean fold change of transcripts in orbicularis oculi compared with quadriceps; the latter was set as 1. Results were analyzed using the Student’s t test: *, P < 0.05; ***, P < 0.0001. (B) MYH13 gene expression was performed by qPCR as described in Materials and methods. Each reaction was performed in triplicate, in pooled muscle samples from five biopsies from different individuals. Expression levels were normalized to ACTN2 expression. Results are expressed as mean fold change of transcripts in orbicularis oculi (OO) compared with quadriceps (QU); the latter were set as 1. Results were analyzed using the Student’s t test: ***, P < 0.0001. The inset shows MyHC protein expression in EOM and orbicularis oculi muscle homogenates; 10 and 30 µg protein were loaded per lane, respectively. Gels and conditions were as described in Materials and methods. (C) Western blot analysis of total SR proteins in human orbicularis oculi and quadriceps muscles. 30 µg of protein was loaded per lane and separated on 6% or 10% SDS-PAGE. Blots were probed with the indicated antibodies. Bar histograms represent the mean (±SEM) band intensity normalized to SRL content. *, P < 0.05. (D) Western blot of human quadriceps, EOM, orbicularis oculi, and mouse heart (positive control) total SR proteins, probed with Cav1.2 antibody. 30 µg of protein was loaded per lane and separated on 6% SDS-PAGE. (E) Western blot of total muscle extracts; 30 µg of protein was loaded per lane, separated on 6% SDS-PAGE, and probed with anti-utrophin (top) and anti-dystrophin (bottom) antibodies. Bar histograms represent the mean (±SEM) band intensity normalized to MyHC content. Student’s t test: *, P < 0.05.

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