Figure 4. Biochemical characterization... | Rockefeller University Press

Figure 4.

$Figure 4. Biochemical characterization of WB-associated WSC. (A) HEX and WSC cofractionate. Organelles were separated on a 17–60% Nycodenz gradient and fractions were probed for various organellar markers. The graph shows the shape of the gradient and total protein distribution across the gradient. Anti-HEX and anti-HA reveal HEX and WSC and antithiolase provides a marker of the peroxisome matrix. The inner mitochondrial membrane protein porin reveals the distribution of mitochondria. (B) WSC is insoluble in Triton X-100. (left) 16 KgP was resuspended in buffer containing the indicated additions and then fractionated into pellet (P) and supernatant (S) fractions by centrifugation at 100,000 g for 45 min. Porin provides a control of an integral membrane protein. (right) An extract obtained from the WSC–GFP–expressing strain received the indicated treatments and was then examined by epifluorescence (WSC–GFP) and bright field (differential interference contrast) microscopy. Bar, 2 μm.$

Biochemical characterization of WB-associated WSC. (A) HEX and WSC cofractionate. Organelles were separated on a 17–60% Nycodenz gradient and fractions were probed for various organellar markers. The graph shows the shape of the gradient and total protein distribution across the gradient. Anti-HEX and anti-HA reveal HEX and WSC and antithiolase provides a marker of the peroxisome matrix. The inner mitochondrial membrane protein porin reveals the distribution of mitochondria. (B) WSC is insoluble in Triton X-100. (left) 16 KgP was resuspended in buffer containing the indicated additions and then fractionated into pellet (P) and supernatant (S) fractions by centrifugation at 100,000 g for 45 min. Porin provides a control of an integral membrane protein. (right) An extract obtained from the WSC–GFP–expressing strain received the indicated treatments and was then examined by epifluorescence (WSC–GFP) and bright field (differential interference contrast) microscopy. Bar, 2 μm.