EMBO/soda-nel huggingface dataset:
Msps Governs Acentrosomal Microtubule Assembly and Reactivation of Quiescent Neural Stem Cells (2021)
Figure 7

Figure 7. Kin-2 and E-cad act downstream of Msps to promote NSC reactivation

A) Larval brains at 24 h ALH from wild-type control (UAS-β-Gal RNAi; UAS-β-Gal RNAi), msps RNAi control (UAS-GFP + UAS-msps RNAi (21982)), and rescued animals (UAS-klp64D+ msps RNAi (21982)) under the control of insc-Gal4 were analyzed for EdU incorporation, and larval brains were labeled with EdU and Dpn.

B) Quantification graph of EdU-negative NSCs per brain lobe for genotypes in (A). n=13 BL for contorl; n=13 BL for msps RNAi; n=20 BL for UAS-klp64D + msps RNAi. ****p<0.0001.

C) Quantification graph of NSC diameter for genotypes in (A). n=246 NSCs for control; n=494 NSCs for msps RNAi; n=457 NSCs for UAS-klp64D + msps RNAi. ****p<0.0001.

D) Larval brains at 24 h ALH from wild-type control (UAS-β-Gal RNAi), RNAi control (UAS-msps RNAi (21982)), and rescued animals (g-kap3 + msps RNAi (21982)) driven by insc-Gal4 were examined for EdU incorporation, and larval brains were stained with EdU and Dpn.

E) Quantification graph of NSCs negative for EdU incorporation per brain lobe for genotypes in (D). n=13 BL for control; n=15 BL for msps RNAi; n=19 BL for g-kap3 + msps RNAi. ****p<0.0001.

F) Quantification graph of NSC diameter for genotypes in (D). n=339 NSCs for control; n=357 NSCs for msps RNAi; n=409 NSCs for g-kap3 + msps RNAi. Error bars represent SD.****p<0.0001.

G) 24 h ALH Larval brains from wild-type control (UAS-β-Gal RNAi; UAS-β-Gal RNAi), RNAi control (UAS-GFP + UAS-msps RNAi (21982)), and rescued animals (UAS-E-cad7 + msps RNAi (21982)) were assessed for EdU incorporation, and larval brains were stained with EdU and Dpn.

H) Quantification graph of NSCs negative for EdU incorporation per brain lobe for genotypes in (G). n=21 BL for control; n=21 BL for msps RNAi; n=22 BL for UAS-E-cad7 + msps RNAi. ****p<0.0001.

I) Quantification graph of NSC diameter for genotypes in (G). n= n=127 NSCs for control; n=230 NSCs for msps RNAi; n=210 NSCs for UAS-E-cad7 + msps RNAi. ****p<0.0001.

J) 24 h ALH Larval brains from wild-type control (UAS-β-Gal RNAi), mutant control (UAS-β-Gal RNAi + klp64Dk5h), and rescued animals (UAS-E-cad7 + klp64Dk5h) were processed for EdU incorporation, and larval brains were probed by with EdU, Dpn, and Mira.

K) Quantification graph of NSCs negative for EdU incorporation per brain lobe for genotypes in (J). n=10 BL for control; n=12 BL for klp64Dk5h; n=10 BL for UAS-E-cad7 + klp64Dk5h. ****p<0.0001; p=0.3006 (ns).

L) A working model. Microtubule arrays in the primary protrusion of Drosophila qNSCs are predominantly acentrosomal with plus-end-out orientation. Msps regulates microtubule growth and orientation in quiescent NSCs. Msps and Kinesin-2 target E-cadherin to the NSC-neuropil contact for NSC reactivation.

Data information: Arrows point at EdU-negative NSCs in A, D, G and J. In (B-C, E-F, H-I, K), data are presented as mean ± SD. In (B-C, E-F, H-I, K), statistical significances were determined by one-way ANOVA with multiple comparisons. Scale bars: 10 µm.

Expended view figure legends
Follistatin-like 1 promotes cardiac fibroblast activation and protects the heart from rupture (2016)
Figure 1

Figure 1. Fstl1 expression is induced in ischemic heart. (A) Detection of Fstl1 protein in ischemic myocardium of WT mice. After MI induction, the heart was harvested at the indicated time points, and separated to ischemic and remote (non-ischemic) areas. Fstl1 protein was detected by western blotting (n=3, each time point). Statistical analysis was performed by ordinary one way ANOVA. Post Hoc test was performed by Dunnett test. (B) Serum Fstl1 was detected by western blotting at the indicated time points. Ponceau S staining of serum reveals equivalent amount of loaded protein (n=4, each time point). Error bars represent mean ± SEM. Statistical analysis was performed by ordinary one way ANOVA. Post Hoc test was performed by Dunnett test.
Follistatin-like 1 promotes cardiac fibroblast activation and protects the heart from rupture (2016)
Figure 2

Figure 2. Fstl1 protein expression is induced in non-cardiomyocyte cells after LAD ligation. (A) DAB immunohistochemical staining of Fstl1 with ischemic heart at day 3 after LAD ligation. Arrows indicate a subset of Fstl1 positive cells. Counter staining was performed with hematoxylin. Scale bar indicates 50μm. (B) Immunofluorescent staining of Fstl1 (green) and sarcomeric actin (red). Fstl1 protein was detected in interstitial areas of the ischemic and border zone but not in cardiomyocytes. Scale bar indicates 100μm. (C) Immunofluorescent staining of Fstl1 (green) and S100a4 (red) in the ischemic area. Arrows indicate Fstl1 and S100a4 double positive cells. Scale bar indicates 20μm. (D) Immunofluorescent staining of Fstl1 (green) and α-smooth muscle actin (red). Arrows indicate Fstl1 and α-SMA double positive cells. Scale bar indicates 20μm. Nuclei were stained by DAPI.
Follistatin-like 1 promotes cardiac fibroblast activation and protects the heart from rupture (2016)
Figure 3

Figure 3. Fstl1 deletion in S100a4cre+ x Fstl1flox+/flox+ mice. Genotypes are indicated as WT for S100a4cre-/- x Fstl1flox+/flox+ mice and cfKO for S100a4cre+/- x Fstl1flox+/flox+. (A) S100a4 expression is induced in the ischemic area (IA). qPCR analysis of mRNA expression of S100a4 in ischemic heart and sham operated heart of WT mice and cfKO mice. Heart samples were harvested at 7 days after the surgery. Statistical analysis was performed by two-way ANOVA. Post Hoc test was performed by Tukey test. Error bars represent mean ± SEM (n=16 and 15 for WT and cfKO sham group, n=15 and 14 for WT and cfKO MI group, respectively). (B) qPCR analysis of mRNA expression of Fstl1 and Tgfβ1 in sham and post-MI heart. Error bars represent mean ± SEM (n=16 and 15 for WT and cfKO sham group, n=15 and 14 for WT and cfKO MI group, respectively). Statistical analysis was performed by two-way ANOVA. Post Hoc test was performed by Tukey test. (C) Western blot analysis of Fstl1 protein expression in ischemic and sham-operated hearts at day 7 after the surgery. Quantified values of Fstl1 protein in WT and cfKO mouse hearts normalized by GAPDH band intensity are shown. Statistical analysis was performed by two-way ANOVA. Post Hoc test was performed by Tukey test. Error bars represent mean ± SEM (n=5 for each sham group and n=6 for each MI group). (D) Fstl1 protein expression in isolated cardiac fibroblasts from cfKO and littermate WT neonatal mice. Cell lysate and its media from cells cultured for 24 hours without FBS were assessed by western blotting. Error bars represent mean ± SEM (n=3 per group). Statistical analysis was performed by unpaired t-test (2-tailed). The experiments were performed twice independently.
Follistatin-like 1 promotes cardiac fibroblast activation and protects the heart from rupture (2016)
Figure 4

Figure 4. Higher mortality due to post-infarction cardiac rupture in S100a4cre+/- x Fstl1flox+/flox+ mice. (A) Mouse survival curve after MI and sham surgery. The mortality of the WT and Fstl1-cfKO mice after MI surgery was 27.1% and 46.7%, respectively. Log-rank (Mantel-Cox) test was used for statistical analysis (n=16 for WT sham, n=15 for Fstl1-cfKO sham, n=59 for WT MI and n=30 for Fstl1-cfKO MI). (B) Representative macroscopic images of cardiac rupture. Left; impending LV rupture found in a cfKO mouse post-MI when sampling. Right; evidence of hemothorax that was diagnosed as cardiac rupture in deceased mice. (C) Representative microscopic images of ruptured heart stained by H&E. Penetrating hematoma due to a tear from the LV endocardium to the epicardium. Left; an image obtained with a low magnification lens. Scale bar indicates 200μm. Right; higher magnification image. Scale bar indicates 100μm. Arrow indicates the tear site. H indicates hematoma.
Follistatin-like 1 promotes cardiac fibroblast activation and protects the heart from rupture (2016)
Figure 5

Figure 5. Impaired ECM protein synthesis in infarcted area of S100a4cre+/- x Fstl1flox+/flox+ mice. (A) Transcript mRNA expression of Col1a1 and Fn1 in sham and post-MI hearts. Hearts were obtained at day 7 after surgery. Error bars represent mean ± SEM (n=13-16 for each group). Statistical analysis was performed by two-way ANOVA. Post Hoc test was performed by Tukey test. (B) Protein expression of collagen I, fibronectin and αSMA in the infarcted and sham hearts at day 7 after surgery, as assessed by western blotting. Quantified values of proteins of interest were normalized relative to the tubulin band intensity. Error bars represent mean ± SEM. Statistical analysis was performed by unpaired t-test (2-tailed) for fibronectin and αSMA and non-parametric t-test (2-tailed) for collagen I (fibronectin and collagen: n=5 per group, αSMA: n=3 and 4 for WT-IA and cfKO-IA, respectively). (C) α-smooth muscle actin-positive myofibroblasts (red) were stained in histological sections from infarcted hearts of WT and cfKO mice. Samples were harvested at day 7 after MI. Counter staining was performed by hematoxylin. Scale bar indicates 100μm for top and middle images, 50µm for bottom images. Error bars represent mean ± SEM. Statistical analysis was performed by Mann-Whitney test (n=16 for WT and n=14 for cfKO). (D) Representative images of Picro Sirius Red staining of infarcted heart at 7 days after MI surgery. Images were taken by standard light (top) and polarized light (bottom) microscopes. Scale bar indicates 100μm. Polarized collagen fibers were sorted to orange-red (middle) and green-yellow (bottom) fibers by Image J software. Error bars represent mean ± SEM. Statistical analysis was performed by unpaired t-test (2-tailed) (n=11 for WT and n=13 for cfKO).
Follistatin-like 1 promotes cardiac fibroblast activation and protects the heart from rupture (2016)
Figure 6

Figure 6. TGF-β1 promotes Fstl1 induction in cardiac fibroblasts. (A) Immunofluorescence staining of Fst1 (green) and α-SMA (red) in NRCFbs at 24 hours after recombinant TGF-β1 (10ng/ml) stimulation. Nuclei were stained by DAPI. Scale bar indicates 50μm. (B) Time course changes of Fstl1 protein expression in NRCFbs after stimulation of recombinant TGF-β1 (10ng/ml). Protein expression of Fstl1 was detected by immunoblotting. Tubulin was used as a loading control (n=3 for each time point). Two independent experiments were performed. (C) Transcript level of Fstl1, S100a4 and Acta2 mRNA in NRCFbs was determined by qPCR analysis. The samples were harvested at 24 hours after stimulation with recombinant TGF-β1 (10ng/ml) or control vehicle. Error bars represent mean ± SEM (Fstl1 and S100a4: n=6 for each group, Acta2: n=3 for each group). Statistical analysis was performed by unpaired t-test (2-tailed) for Fstl1 and S100a4, and non-parametric unpaired t-test (2-tailed) for Acta2. Two independent experiments were performed.
Follistatin-like 1 promotes cardiac fibroblast activation and protects the heart from rupture (2016)
Figure 7

Figure 7. Fstl1 promotes the phosphorylation of ERK1/2 and cardiac fibroblast migration. (A) NRCFbs at passage 1 were stimulated with recombinant Fstl1 (50ng/ml) or vehicle after cultured in serum-reduced conditions (FBS 0.5%) for 24 hours. The samples were harvested at the indicated time points after stimulation. The expression of ERK1/2 and tubulin were detected by immunoblotting. Error bars represent mean ± SEM. Statistical analysis was performed by one-way ANOVA and Tukey multi comparison test (n=3 for each time point). Three independent experiments were performed. (B) Fstl1 induced phosphorylation of ERK1/2 was ablated by pretreatment with MEK inhibitor PD98059. NRCFbs were treated by PD98059 (5nM) for 30min and then stimulated with recombinant Fstl1 protein (50ng/ml) for 15min. Error bars represent mean ± SEM. Statistical analysis was performed by one-way ANOVA and Tukey multi comparison test (n=3 for each time point). Two independent experiments were performed. (C) Representative images of NRCFbs scratch assay (left) and quantified cell migration (right). NRCFbs were transfected by Fstl1 siRNA or siRNA non-targeting negative control for 12 hours followed by culturing in 0.5% FBS media for 24 hours. The confluent cell sheet was scratched and cell migration was assessed at 6 hours after the scratch. Scale bar indicates 100µm. Error bars represent mean ± SEM (n=10 for each group). Statistical analysis was performed by unpaired t-test (2-tailed). Two independent experiments were performed. (D) Fstl1-stimulation of cell migration was reversed by PD98059. Endogenous Fstl1 in NRCFbs was ablated by Fstl1 siRNA. Serum-deprived NRCFbs were treated with PD98059 (5nM) for 30 min and then stimulated by Fstl1 (50ng/ml) or vehicle. Cell migration was assessed at 6 hours after Fstl1 stimulation. Error bars represent mean ± SEM (n=7-9 for each group). Statistical analysis was performed by Kruskal-Wallis test and Dunnett’s T3 test. Two independent experiments were performed. (E) Morphological changes of NRCFbs after recombinant Fstl1stimulation was assessed by immunocytochemistry. 24 hours serum starved NRCFbs was stimulated by Fstl1 (50ng/ml) or vehicle for 3 hours. PD98059 (5nM) or DMSO was added 30min prior to Fstl1 stimulation. Cells were stained with Alexa Fluor 488-conjugated phalloidin antibody and nuclei were stained by DAPI. Arrow shows lamellipodium of the cell. Scale bar indicates 50μm.
Follistatin-like 1 promotes cardiac fibroblast activation and protects the heart from rupture (2016)
Figure 8

Figure 8. Fstl1 promotes cardiac fibroblasts proliferation via the ERK1/2 dependent pathway. (A, B) Fibroblast proliferation was assessed by Edu incorporation assay. Endogenous Fstl1 was ablated by siRNA. NRCFbs were cultured in 0.5% FBS condition for 48 hours to synchronize the cell cycle. EdU (10μM as the final concentration) was added into media at 4 hours before harvest. Error bars represent mean ± SEM (n=5, each group). Statistical analysis was performed by unpaired t-test (2-tailed). Two independent experiments were performed. (B) Effect of exogenous Fstl1 on cardiac fibroblast proliferation. Cells were cultured in FBS 0.5% media for 24 hours. Recombinant Fstl1 (50ng/ml) or vehicle were added to 2% FBS-containing media and cultured for 48 hours. EdU was added into media at 4 hours before harvest. Error bars represent mean ± SEM (n=7, each group). Statistical analysis was performed by unpaired t-test (2-tailed). Two independent experiments were performed. (C) Fstl1 promoted fibroblasts proliferation was diminished by PD98059. PD98059 (5nM) was added 30 min prior to recombinant Fstl1 stimulation (50ng/ml). The cells were cultured for 48 hours, and EdU was added into media at 4 hours before harvest. Error bars represent mean ± SEM (n=4-10, per treatment group). Statistical analysis was performed by one-way ANOVA and Tukey multi comparison test. Two independent experiments were performed. (D) Schema of the role of Fstl1 in infarct repair. Fstl1 is upregulated by TGF-β1 in infarcted heart and it contributes to the proliferation and migration of fibroblasts into the infarcted site and border zone. These actions increase the number of myofibroblasts (MyoFB) in the infarcted lesion. In turn, fibroblasts and myofibroblasts synthesize ECM components including collagens and fibronectin, and protect the infarcted heart from rupture.
Phosphopeptide binding by Sld3 links Dbf4-dependent kinase to MCM replicative helicase activation (2016)
Figure 1

Figure 1. DDK promotes Sld3-dependent Cdc45 recruitment. (a) Reaction scheme for Sld3/7/Cdc45 recruitment experiments. DNA substrate was bound to beads and reaction mixtures were removed after each step. (b, c) Immunoblots of recruitment reactions performed as described in (a), with the indicated proteins omitted. In (c), a mid reaction wash in high salt (0.5 M NaCl) buffer (HSW) was included following DDK phosphorylation as indicated. (d) Recruitment reaction performed as in (a), using wild type (wt) or a Cdc45-binding mutant (3E1) of Sld3. (e) Schematic showing position of Cdc45-interacting region in Sld3. Alignments were generated in Jalview using Clustal. Sld3 from various fungal species is included (S.c., Saccharomyces cerevisiae, S.m., Saccharomyces mikatae, S.b., Saccharomyces bayanus, S.ca., Saccharomyces castellii, S.ku., Saccharomyces kudriavzevii). Residue numbers correspond to S. cerevisiae Sld3. Residues were substituted for Glu. as indicated. (f) Wild type or mutant FLAG-Sld3 was added to an S-phase protein extract and tested for interaction with Cdc45. Immunoprecipitated proteins were analysed by immunoblot. (g) Reaction scheme for in vitro replication reactions. Reaction mixtures were removed after each step. (h) Reaction performed as in (g). Nascent DNA was separated in a 0.7% alkaline agarose gel in this and all subsequent replication reactions.
Phosphopeptide binding by Sld3 links Dbf4-dependent kinase to MCM replicative helicase activation (2016)
Figure 2

Figure 2. Mapping the MCM binding activity in Sld3/7. (a) Reaction performed as in Fig. 1a, except Cdc45 was omitted. (b) Reaction performed as in Fig. 1g, with the indicated proteins omitted. (c) Schematic depicting MCM-binding by Sld3 fragments. Two essential CDK phosphorylation sites are shown by vertical bars. The Sld3-Treslin Domain (S.T.D.) is shown as a black box. Also see Fig. EV1 (d) Schematic showing position of MCM-interacting region in Sld3. Alignments were generated as in Fig. 1e. Residues were substituted for Glu. as indicated. (e) Reaction performed as in (a), except Sld3/7 was incubated with Rad53 before addition to reactions. ATP was omitted from this pre-incubation as indicated. (f) Sld3 recruitment reaction performed as in (a). Sld7 was omitted from this experiment.
Phosphopeptide binding by Sld3 links Dbf4-dependent kinase to MCM replicative helicase activation (2016)
Figure 3

Figure 3. MCM binding is an essential function of Sld3. (a) Reaction scheme for complete firing factor recruitment reaction conducted on ARS305 linear DNA. (b) Recruitment reaction performed as described in (a). (c) Sld3/7 recruitment reaction conducted as in Fig. 2a. (d) Replication reaction performed as in Fig. 1g. (e) Representative tetrads from dissection of SLD3+/sld3-6E heterozygotes.
Phosphopeptide binding by Sld3 links Dbf4-dependent kinase to MCM replicative helicase activation (2016)
Figure 4

Figure 4. Sld3 binds to phosphorylated Mcm4/6. (a) Reaction scheme to examine interactions of Sld3/7 with individual Mcm subunits. Phosphorylated MCM bound to DNA was incubated in buffer containing 0.5 M NaCl to cause dissociation into individual subunits. Interactions with Sld3/7-coupled beads were then examined. (b, c, D) Immunoblot of reactions performed as described in (a). In (B), either reconstituted Sld3/7 complex (lanes 6 and 7) or Sld3 alone (lanes 8 and 9) was coupled to beads. In (C), MCM was treated with Lambda Phosphatase after dissociation from DNA (lanes 3 and 7). In (D), Sld7 was omitted. (E) 30-residue phosphorylated Mcm6 peptides were bound to streptavidin-coated beads and used as bait to pulldown Sld3. Peptide-coupled beads were dephosphorylated with Lambda Phosphatase as indicated. (F) Reaction conducted as in (a), using wild type (wt) or the indicated S/T-A substitution mutant versions of Mcm4 and Mcm6. Positions of mutations in Mcm6 are summarised in Fig. EV5c.
Phosphopeptide binding by Sld3 links Dbf4-dependent kinase to MCM replicative helicase activation (2016)
Figure 5

Figure 5. Sld3 binding mutants in Mcm4/6 are defective in replication. (a) Sld3/7 recruitment reaction performed as in Fig. 2a. (b) Replication reaction performed as in Fig. 1g. Experiments in (a) and (b) used MCM containing wild type (wt) or the indicated S/T-A substitution mutant versions of Mcm4 and Mcm6. (c) Quantification of (b), performed using ImageQuant software (GE Healthcare). (d) Representative tetrads from diploids heterozygous for mcm4-25A and mcm6ΔN+11A.
Phosphopeptide binding by Sld3 links Dbf4-dependent kinase to MCM replicative helicase activation (2016)
Figure 7

Figure 7. DDK-independent replication. (a, b) Recruitment reactions performed as in Fig. 2a. Sld7 was omitted in (b). (c) Recruitment reaction conducted as in Fig. 3a, with washes performed at 0.3 M KCl. (d) Replication reaction performed as in Fig. 1g. Experiments in (a)-(d) used MCM containing wild type (wt) or S/T-D substitution mutants of Mcm4 and Mcm6 as indicated. In (d), MCM loading was equalised across samples by using 5-fold more mutant than wild type MCM protein, as is shown in Fig. EV6c. (e) Model for Sld3/7-dependent Cdc45 recruitment to the MCM double hexamer. Sld3/7 may promote intra- (black arrows) or inter- (red arrows) hexamer Cdc45 recruitment. See text for full details of model in (e).
Bacterial killing by complement requires membrane attack complex formation via surface-bound C5 convertases (2018)
Figure 1

Figure 1. MAC in serum perturbs both the outer and inner membrane of Gram-negative bacteria.

(A) Schematic representation of engineered perimCherry/cytoGFP E. coli cells that express mCherry in the periplasmic space (between the outer and inner membrane) and GFP in the cytosol.

(B) Structured illumination microscopy image of perimCherry/cytoGFP E. coli confirming localization of mCherry (red) in the periplasm and GFP (green) in the cytosol. Scale bar = 3 µm.

(C) Outer membrane damage (mCherry intensity) and inner membrane damage (% Sytox positive) of perimCherry/cytoGFP E. coli bacteria exposed to (different concentrations of) human serum. Inner membrane damage correlates with killing (samples where bacteria are killed are indicated with gray shadings and a cross, see CFU data in Fig EV1b).

D, E (D) Serum-induced inner membrane damage (% Sytox positive) and (E) killing (CFU/ml) of different Gram-negative strains depends on MAC components C5 and C8, but not on lysozyme (10% serum). Dotted line represents the detection limit of the assay.

Data information: (D, E) The cfu/ml (Fig. 1E, 2C) and sytox measurements (Fig. 1D, 2E) of "Buffer", "Serum", "ΔC5", "ΔC8" "Δlysozym" and "C5b6MAC" were all generated from the same experiment. (C-E) Data represent mean ± SD of 3 independent experiments. (D, E) Statistical analysis was done using a ratio paired two-tailed t-test and displayed only when significant as * p≤0.05, ** p≤0.01, *** p≤0.001 or **** p≤0.0001.
Bacterial killing by complement requires membrane attack complex formation via surface-bound C5 convertases (2018)
Figure 3

Figure 3. Reconstituting bactericidal MAC assembly via surface-bound C5 convertases.

(A) Schematic overview for Conv-MAC formation. Bacteria were labeled with C5 convertases by pre-incubation with C5-deficient serum (Fig EV3). Following a washing step (@), convertase-labeled bacteria were incubated with uncleaved C5, C6, C7, C8 and C9 (termed 'Conv-MAC').

(B) Bacterial viability of convertase-labeled bacterial strains exposed to buffer (Conv) or C5-C9 (Conv-MAC).

(C) Bacterial viability of convertase-labeled E. coli MG1655 exposed to a concentration range of C5 in the presence of 100 nM C6, 100 nM C7, 20 nM C8 and 100 nM C9. 'Ctrl' indicates bacteria that are pretreated with heat-inactivated ΔC5 serum. Dotted line represents the detection limit of the assay.

(D) Bacterial viability of convertase labeled E. coli MG1655 exposed to C5-C9 or conditions lacking one MAC component. As an extra control, convertase formation was blocked during ΔC5 serum incubation by adding 5 µM compstatin.

(E) Bacterial viability of E. coli MG1655 exposed to FB depleted serum in the presence of 20 µg/ml OmCI (to deposit C4b and C3b without Bb). After washing, bacteria were exposed to C5-C9 in the presence or absence of C1 and C2 (to generate classical pathway C5 convertases, C4b2aC3b).

Data information: (B-E) Data represent mean ± SD of 3 independent experiments. (B, D, E) Statistical analysis was done using a ratio paired two-tailed t-test and displayed only when significant as * p≤0.05, ** p≤0.01, *** p≤0.001 or **** p≤0.0001.