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. Author manuscript; available in PMC: 2022 Aug 9.
Published in final edited form as: Nat Cell Biol. 2020 Jan 6;22(1):74–86. doi: 10.1038/s41556-019-0441-z

Figure 4. Golgi and post-Golgi transport of PC-I is rhythmic.

Figure 4

(A) Immunofluorescence analysis of collagen chaperone HSP47, and ERGIC and the cis-Golgi (detected by marker GM130) in synchronised MEFs showed gradual co-localisation of HSP47 with GM130 (see insets), n=5. Quantification in Figure S6D. (B) Immunofluorescence analysis shows that PDE4D depletion in MEFs using siRNAs (Pde4d KD) blocked collagen-I secretion into the matrix, which was unaffected in control cells, n=3. (C) Immunofluorescence analysis of HSP47 in Pde4d KD cells show accumulation of HSP47 in GM130 compared to control MEFs, n=5. (D) Immunofluorescence analysis of synchronised iTTFs shows collagen-I secretion (white arrows) was inhibited in CRISPR-mediated Vps33b KO cells, where collagen-I was retained in the ER (PDI staining) (yellow arrows). N=4 biologically independent experiments. (E) Immunofluorescence analysis of synchronised MEFs also shows increased retention of collagen-I occurs in the Golgi (GM130 staining) when Vps33b was knocked out (see insets). N=2 biologically independent experiments. Bars 10 μm.