Figure S6.
Graph A (I E M Correction Factor (My B P C)): The vertical axis represents the Correction Factor (unitless), ranging from 1.00 to 1.15 with increments of 0.05, while the horizontal axis represents Sarcomere Length in micrometers, ranging from 1800 to 2600 with increments of 200. A linear regression line follows the equation y equals minus 0.0001299x plus 1.457 across the black data points. Graph B (M8-M10 versus S L): The vertical axis shows the Distance from M-line in nanometers, ranging from 0 to 80 with increments of 20, and the horizontal axis shows Sarcomere Length in nanometers, ranging from 1800 to 2800 with increments of 200. W T (gray circles) and Ttn Delta A164-167 (blue circles) data points cluster horizontally around 40 nanometers. Graph C (A168-170 versus S L): The vertical axis measures Distance from M-line in nanometers, ranging from 0 to 200 with increments of 50, while the horizontal axis measures Sarcomere Length in nanometers, ranging from 1800 to 2800 with increments of 200. Three distinct populations are visible: W T alpha (top gray) at approximately 120 nanometers, W T beta (middle gray) at approximately 80 nanometers, and Ttn Delta A164-167 (blue) at approximately 70 nanometers. Graph D (Ti102/M I R versus S L): The vertical axis represents Distance from M-line in nanometers, ranging from 400 to 1200 with increments of 200, and the horizontal axis represents Sarcomere Length in nanometers, ranging from 1500 to 2500 with increments of 500. Both W T (gray) and Ttn Delta A164-167 (blue) show a positive linear slope, with vertical dashed lines marking a specific range between 1700 and 2300 nanometers. Graph E (A77-78 versus S L): The vertical axis measures the Distance from M-line in nanometers, ranging from 300 to 600 with increments of 100, while the horizontal axis represents Sarcomere Length in nanometers, ranging from 1500 to 3000 with increments of 500. Vertical dotted lines at approximately 1700 and 2300 nanometers highlight a specific analysis range where W T data (gray) appears slightly higher than the mutant data (blue). Graph F (cMyB P C stripe 1 versus S L): The vertical axis represents Distance from M-line in nanometers, ranging from 100 to 200 with increments of 20, and the horizontal axis represents Sarcomere length in nanometers, ranging from 1800 to 2800 with increments of 200. The W T data points (gray) show a distribution centered around 165 nanometers, which is significantly higher than the mutant distribution (blue) centered near 125 nanometers. Graph G (cMy B P C stripe 9 versus S L): The vertical axis measures Distance from M-line in nanometers, ranging from 400 to 600 with increments of 50, while the horizontal axis represents Sarcomere length in nanometers, ranging from 1800 to 2800 with increments of 200. Consistent with panel F, the W T data (gray) sit at a higher position (approximately 510) compared to the mutant data (blue), which clusters around 465 nanometers. All values are approximate.
Shrinkage correction and epitope distance vs. sarcomere length relationship. (A) Correction factor calculated for each image stained with cMyBP-C antibody is plotted against the average sarcomere length of the sarcomeres in that image to generate a standard curve whose equation can be used to calculate the correction factor for an image not stained with cMyBP-C antibody based on the average length of the sarcomeres in that image. (B–G). Epitope distance from M-band vs. sarcomere length for: B. M8–M10, C. A168–170, D. A77–78, E. Ti102/MIR, F. cMyBP-C stripe 1, and G. cMyBP-C stripe 9. Data in manuscript are restricted to SL = 1,700–2,300 nm (indicated by dotted vertical lines) for A77–78 and Ti102/MIR.