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Figure 5


Fig. 5. In the absence of L chains, µ{Delta}CH1 condense primarily in tubular vesicles and saccules enriched in ERGIC-53. (a-f) H[µ{Delta}CH1] cells were cultured with (a,b) or without tetracycline for 3 (c,d) or 7 days (e,f) and stained with antibodies against ERGIC-53 (a,c,e in green), GM130 (b,d,f; green) or µ (c-f; red). Note that ERGIC-53 abandons its canonical distribution (a) to co-localize with µ{Delta}CH1-containing sRB upon tetracycline removal (panels c,e yellow staining). µ{Delta}CH1 accumulates close to GM130, but does not overlap with it (d,f); bar, 5 µm. (g,h) After 7 days of culture without tetracycline, cells were processed for cryo-EM with anti-µ (g) or anti-ERGIC-53 (h, G1/93) antibodies and revealed with protein-A coupled to 15 nm gold particles. µ{Delta}CH1 chains accumulate in saccules enriched in ERGIC-53. Bars, 100 nm. (i,j) H[µ{Delta}CH1] cells were kept for 7 days without tetracycline and processed for resin embedding and EM. Continuity between rough ER and µ{Delta}CH1-containing sRB are evident in j (see arrow; the arrowheads point to regions rich in ribosomes). Bars, 100 nm.