Search the tables and observations reproduced from the product literature guide. Rows may represent measurements, recipes or experimental context; they are not all independent experiments.
Parameter: Loading ratio | Reported procedure / limitation: MK16:mRNA = 12.5:1 by mass. This applies to MK16 alone and is not N/P or total lipid:mRNA. | Source: Prior main-article review; Supplementary Fig.6a documents MK6E formulation selection
Parameter: Recipe lineage | Reported procedure / limitation: The formulation screen first varied MK6 molar parts 20, 40 and 60 while DOPE/cholesterol/PEG-lipid remained 30/40/0.75; then MK6:mRNA mass ratios 7.5, 10, 12.5 and 15 were compared. MK16 is the later structurally optimized lipid, not a synonym for MK6 or MK6E. | Source: Supplementary Fig.6a; main Fig.3
Parameter: Mixing and purification | Reported procedure / limitation: The earlier article review notes that preparation cross-references a prior method. Exact buffer composition/pH, flow settings and purification parameters for a fully reproducible MK16 preparation have not been recovered here. | Source: Prior main Methods review; newly recovered SI does not supply a complete standalone formulation protocol
Parameter: Particle measurements | Reported procedure / limitation: Size and PDI, encapsulation, surface charge and cryo-EM are separately reported for MK16. Use MK16 Fig.S9 measurements rather than MK6 Fig.S5 values. | Source: Supplementary Fig.9 and deposited Supplementary Data, sheet Fig.S9
Measurement / batch: Hydrodynamic diameter | Reported result: 137.0 ± 4.1 nm; recomputed from 133.1, 141.2 and 136.8 nm. Mean±sample SD, n=3. | Evidence context: Supplementary Fig.9a; Supplementary Data, Fig.S9!B2:B4
Measurement / batch: PDI | Reported result: 0.108 ± 0.014; recomputed mean±sample SD from 0.099, 0.101 and 0.125, n=3. | Evidence context: Supplementary Fig.9a; Supplementary Data, Fig.S9!C2:C4
Measurement / batch: mRNA encapsulation | Reported result: 84.84 ± 1.50%; mean±sample SD, n=3. | Evidence context: Supplementary Fig.9b; Supplementary Data, Fig.S9!B7:B9
Measurement / batch: Zeta potential | Reported result: +5.87 ± 1.98 mV; mean±sample SD, n=3. | Evidence context: Supplementary Fig.9b; Supplementary Data, Fig.S9!C7:C9
Measurement / batch: Apparent particle pKa | Reported result: 6.86 by the TNS assay; explicitly printed in the fitted-curve inset. | Evidence context: Supplementary Fig.6d, SI printed p.28 / PDF p.29
Experiment and dose: Brain reporter expression Mouse; FLuc mRNA; intravenous; 0.5 mg/kg (dose/time from prior main-article review) | Readout and comparator: Readout: 6 h Comparator: MC3 and MK6E formulations | Result and source: MK16 brain signal was approximately 8.3-fold MC3 and 1.7-fold MK6E; deposited Fig.3d has n=3 per group. Separate SI8 comparator experiment gives approximately 7.4-fold ALC-0315 and 6.5-fold SM-102. Fig.3d; Source Data Fig.3, sheet Fig.3d; Supplementary Fig.8 / Data sheet Fig.S8
Experiment and dose: Brain cell-type expression Mouse; GFP mRNA; intravenous; 1 mg/kg (dose/time from prior main-article review) | Readout and comparator: Readout: 12 h Comparator: PBS and MC3 at 0.5 or 1 mg/kg | Result and source: GFP-positive neurons 7.36±0.78%, astrocytes 9.71±0.71%, microglia 2.85±0.44%, and brain capillary endothelial cells 9.18±0.80%. n=3; mean±sample SD recalculated from source data. These are percentages within the respective cell populations. Fig.3f; Source Data Fig.3, Fig.3f!J8:L11
Experiment and dose: Fluorescent RNA tissue distribution MK16 carrying Alexa-Fluor-647-labelled RNA; intravenous; 0.5 mg/kg in the prior main-article review | Readout and comparator: Readout: 1 h Comparator: Six measured organs | Result and source: Brain accounted for 15.33±0.38% of the summed measured-organ fluorescence, n=3. This is not percent injected dose and is not the percentage of functional mRNA expression in the brain. Supplementary Fig.10b; Supplementary Data, Fig.S10b!A2:A4
Experiment and dose: Functional reporter recombination Ai14 mice; MK16-Cre mRNA; one versus three intravenous administrations | Readout and comparator: Readout: Endpoint timing and dose require the complete main Fig.4 protocol Comparator: MC3-Cre mRNA and PBS | Result and source: Deposited Fig.4i shows neuronal tdTomato positivity 8.28±0.65% after one and 16.73±2.15% after three administrations; astrocytes 10.10±1.06% and 19.20±2.98%, respectively (n=3). Recombination accumulates and is not equivalent to instantaneous GFP transfection efficiency. Publisher Fig.4i and caption; Source Data Fig.4, sheet Fig.4i, G8:L9; SI Discussion and Fig.13
Experiment and dose: Ex vivo human cortical tissue Adult human cortical tissue exposed to MK16 carrying ΔFosb mRNA | Readout and comparator: Readout: Dose and exposure duration not re-recovered; do not infer from mouse experiments Comparator: PBS | Result and source: Source Fig.5e records ΔFOSB-positive neurons 4.00±0.99%, astrocytes 6.49±2.33%, and microglia 2.38±0.94%, n=4 tissue slices per group (not a verified four-donor cohort). This is ex vivo expression and does not demonstrate intravenous delivery across a human BBB. Versus PBS: neuron P=0.0004, astrocyte P=0.0020, microglia P=0.0065 by two-tailed Student’s t-test. Publisher Fig.5e and caption; SI Discussion pp.3–4; Source Data Fig.5, Fig.5e!H8:K10
Experiment and dose: Glioblastoma efficacy Orthotopic U118MG mouse model; Pten mRNA; intravenous 1 mg/kg; three treatments on days 10,13,16 | Readout and comparator: Readout: Tumour imaging and survival follow-up Comparator: PBS, MK16-control mRNA; separate MC3-Pten comparison | Result and source: The article reports suppression of tumour growth and approximately 70% survival beyond 120 days for MK16-Pten. The separately deposited SI19 comparison with MC3 uses n=10/group and 1 mg/kg. Exact raw survival counts should not be merged across the two experiments. Main Fig.6 caption gives n=9 PBS, n=9 control mRNA and n=10 Pten mRNA. Publisher Fig.6a,d and caption (fresh visual review); Supplementary Fig.19; Source Data Fig.6
Experiment and dose: In vitro BBB transport mechanism bEnd.3 monolayer, 0.4-µm Transwell, 1×10^5 cells/well; culture 7–9 d; TEER >60 Ω·cm²; Alexa-Fluor-647 RNA | Readout and comparator: Readout: Lower-chamber fluorescence at 12 h Comparator: MK16 alone; EIPA 10 µM, MβCD 4.5 mM, CPZ 5 µg/mL or MK-0752 10 nM pretreatment | Result and source: Transport decreased with MβCD and MK-0752; SI12 source data show relatively little change with EIPA/CPZ. The inhibition experiment supports pathway involvement, not an exclusive transport mechanism. SI Methods 2.3, printed p.21; Supplementary Fig.12c and Fig.S12 source data
Experiment and dose: Functional delivery after Transwell crossing bEnd.3 upper chamber and N2a lower chamber; N2a seeded at 1×10^5/well one day earlier; MK16-FLuc mRNA; MβCD 4.5 mM or nirogacestat 10 nM pretreatment | Readout and comparator: Readout: 18 h; Bright-Glo readout after 5 min Comparator: MK16 without inhibitor and PBS | Result and source: Inhibitor-sensitive luciferase expression in both compartments supports transport followed by functional RNA delivery. Dose not specified in the recovered SI method. SI Methods 2.3; main Fig.3i source data and Supplementary Fig.12d
Experiment and dose: In vivo transport inhibition Mice pretreated with MβCD 100 mg/kg or nirogacestat 30 mg/kg 1 h before labelled-RNA MK16 BLNPs | Readout and comparator: Readout: Brain IVIS at 1 h after BLNP administration Comparator: MK16 without inhibitor | Result and source: Both pretreatments reduced brain fluorescence. The inhibitors affect biological pathways and this experiment alone does not establish molecular specificity. SI Methods 2.3; Source Data Fig.3, Fig.3j
Experiment and dose: Single-dose systemic tolerability MK16-FLuc mRNA versus MC3-FLuc mRNA; intravenous 1 mg/kg | Readout and comparator: Readout: 6,24,48 h Comparator: PBS and MC3 | Result and source: 32-plex cytokine/chemokine analysis, ALT, AST, BUN and H&E were assessed. The authors report transient responses with recovery and no detected hepatic/renal injury under these tested conditions; n=3, not a chronic or human safety assessment. SI Methods 2.4, pp.21–22; Supplementary Figs.14–16
Experiment and dose: Repeated-dose tolerability Ai14 Cre-reporter study; single or triple systemic administrations | Readout and comparator: Readout: 5 d after final administration Comparator: PBS and MC3 | Result and source: Cytokines, ALT/AST/BUN and histology were evaluated; Supplementary Fig.17 marks group comparisons nonsignificant (n=3). This later time point does not exclude earlier transient responses. SI Methods 2.4; Supplementary Fig.17, printed pp.40–41