Journal: Cells

Article Title: Post-ER Stress Biogenesis of Golgi Is Governed by Giantin

doi: 10.3390/cells8121631

Figure Lengend Snippet: The overlap of giantin and Rab6a during Golgi biogenesis. ( A ) Confocal immunofluorescence images of Rab6a in HeLa cells after 60 min of BFA-WO, pretreated with scramble, giantin, GM130, or GRASP65 siRNAs. All confocal images acquired with the same imaging parameters; bars, 10 μm. ( B ) Quantification of cells with membranous Rab6a in cells presented in ( A ); n = 90 cells from three independent experiments, results are expressed as a mean ± SD; * p < 0.001. ( C ) Giantin immunostaining in DMSO- and BFA-treated HeLa cells. ( D ) Giantin immunostaining in HeLa cells after 60 min of BFA-WO, transfected with scramble, Rab6a siRNAs, and dominant-negative (GDP-bound) Rab6a(T27N). ( E ) Rab6a W-B of lysates of HeLa cells treated with corresponding siRNAs; β-actin was a loading control. ( F ) Quantifications of cells with perinuclear Golgi in cells from ( C , D ); n = 90 cells from three independent experiments, results expressed as a mean ± SD; *, p < 0.001.

Article Snippet: PCMV-intron myc Rab6 T27N was a gift from Terry Hebert (Addgene, Cambridge, MA, USA, plasmid # 46782) [ ].

Techniques: Immunofluorescence, Imaging, Immunostaining, Transfection, Dominant Negative Mutation, Control

Journal: bioRxiv

Article Title: Golgi Renaissance: the pivotal role of the largest Golgi protein giantin

doi: 10.1101/296517

Figure Lengend Snippet: ( A ) Confocal immunofluorescence images of giantin and ASGP-R in VA-13 cells: control, EtOH-treated cells, EtOH-treated cells and transfected with scramble, giantin, NMIIB, Rab6a siRNAs, and dominant negative (GDP-bound) Rab6a(T27N) followed by recovery. White boxes are enlarged pictures of ASGP-R presented at the right side. Arrowheads indicate ASPR-R punctae distributed at the periphery of cells. All confocal images acquired with same imaging parameters; bars, 10 μm. ( B ) Quantification of cells with compact Golgi in cells presented in A; n = 90 cells from three independent experiments, results expressed as a mean ± SD; *, p<0.001. ( C ) Giantin, NMIIB, and Rab6a W-B of lysates of VA-13 cells treated with corresponding siRNAs; β-actin was a loading control. ( D ) Transferrin, PIGR, and ASGP-R W-B of plasma membrane fractions isolated from VA-13 cells presented in A; samples were normalized to E-cadherin. ( E ) SNA lectin W-B of ASGPR-IP from the plasma membrane fractions isolated from VA-13 cells: control, EtOH-treated, and recovered from EtOH in absence or presence of giantin siRNAs. The input was normalized to the E-cadherin.

Article Snippet: PCMV-intron myc Rab6 T27N plasmid was a gift from Terry Hebert (Addgene plasmid # 46782)( ).

Techniques: Immunofluorescence, Transfection, Dominant Negative Mutation, Imaging, Isolation