MG patients studied in the validation cohort.

<p>M = male; F = female; No  =  No treatment/No thymectomy; Cort  =  corticosteroids; Other immunosuppressors =  OI; Yes  =  Thymectomy.</p

Unsupervised HeatMap showing deltaCt values of the miRNA in differential levels between MG patients and controls in the discovery cohort.

<p>Delta Ct value is the difference between Ct value of the target miRNA minus Ct value of the reference small RNAs. The scale at top indicates high delta Ct values (in red shades) and low delta Ct values (in green shades). All miRNAs with a p adjusted value <0.05 are shown. Early onset MG (green), late onset MG (red), thymoma MG (blue) and healthy controls (purple).</p

Effect of thymectomy on miRNAs levels in the validation cohort.

<p>YES  =  thymectomyzed MG patient; NO  =  not thymectomyzed MG patient.</p

miR15b, miR122, miR140-3p, miR185, miR192, miR20b and miR885-5p show low levels in sera of MG patients.

<p>No differences were found for miR375. Graphs show relative quantification of the miRNAs in the 3; EOMG  =  early onset MG; late onset MG; thymoma  =  thymoma MG; CTRL  =  healthy controls; *P<0.05; **P<0.01.</p

Subjects analyzed in the discovery cohort.

<p>M = male; F =  female.</p

Effect of immunosuppressors treatment on miRNAs levels in the validation cohort.

<p>NO  =  non-treated; Cort  =  patient treated with steroids; Cort + OI  =  patient treated with steroids + other immunosuppressor agent; OI  =  patient treated with immunosuppressor agents other than steroids.</p

Molecular effects of <i>c.2310_2311dupCC</i> mutation and GP isoforms expression in muscle cells.

<p>(A) Electropherogram of <i>PYGM</i> exon 18 in control C1 (top) and patient P1 (bottom). The square shows the <i>c.2310_2311dupCC</i> mutation. Exonic nucleotides are underlined in black; intronic nucleotides are underlined in grey. Encoded amino acids are indicated (including wild type Arg, and mutant Gly, encoded by codons formed in an exon-exon junction). (B) Immunoblotting showing muscle GP and α-tubulin bands in muscle biopsy homogenates from control C1 and patient P1. (C) Relative contribution of each gene (<i>PYGM</i>, <i>PYGB</i> and <i>PYGL</i>), to the total amount of GP mRNA in undifferentiated and 12 day differentiated cultured muscle cells. Bars represent the result of a single experiment for undifferentiated cells, or mean ± SD of two independent experiments for 12 days differentiated cells. Percentages were calculated as [<i>PYG(x)</i> mRNA x 100/(<i>PYGB</i> mRNA + <i>PYGL</i> mRNA + <i>PYGM</i> mRNA)], using values normalized for the <i>PPIA</i> mRNA. In C1 and C2 skeletal muscle (not shown), <i>PYGB</i> mRNA and <i>PYGL</i> mRNA were negligible (<0.5%).</p

Subjects' information.

<p>*Age at the time the skeletal muscle biopsy was collected. GenBank reference sequence was NP_005600.1. Expression values were calculated considering 100% the mean results obtained in the controls (C1, C2).</p

mRNA quantification of GP genes at 0, 7 and 12 days of differentiation.

<p>Results from a single experiment (before differentiation; C1, P1 and P2), or mean of two independent experiments (7-day and 12-day differentiation, C1, C2, P1 and P2) are depicted.</p

Glycogen phosphorylase activity ratio (active/total), glycogen synthase activity ratio (active/total) and glycogen content.

<p>The results represent values for controls (C1, C2) and patients (P1, P2), after 7 days of differentiation. Values are mean ± SEM. The significance of the difference versus controls is: *p<0.01 and **p<0.001. Grey bars represent controls and black bars patients.</p