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  • Abstract
  • Authors
  • Analysis Overview
    • Quality control
  • Licenses
  • Citations
  • Version Information

Last updated: 2024-08-09

Checks: 7 0

Knit directory: paed-inflammation-CITEseq/

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Abstract

Authors

Analysis Overview

Quality control

  1. Empty droplet removal and demultiplexing

    1. Batch 0
    2. Batch 1
    3. Batch 2
    4. Batch 3
    5. Batch 4
    6. Batch 5
    7. Batch 6
  2. Quality control

  3. Identify doublets

  4. Filter doublets

  5. Remove ambient contamination

  6. Annotate using Human Lung Cell Atlas v2.0

    1. Without ambient removal
    2. With ambient removal
  7. Integrate and cluster T-cells

    1. Without ambient removal
    2. With ambient removal
  8. Integrate and cluster other cells

    1. Without ambient removal
    2. With ambient removal
  9. Integrate and cluster macrophage cells

    1. Without ambient removal
    2. With ambient removal
  10. Macrophage annotations

    1. Without ambient removal
    2. With ambient removal
  11. T/NK cell annotations

    1. With ambient removal
  12. Other annotations

    1. With ambient removal
  13. Differential expression analysis within macrophage sub-types

    1. Alveolar macro. CF modifiers & severity
    2. Alveolar macro. CF±modifiers vs non-CF control
    3. Monocyte-derived macro. CF modifiers & severity
    4. Monocyte-derived macro. CF±modifiers vs non-CF control
  14. Cell type proportions

    1. Broad labels
    2. Fine labels
    3. Macrophage labels

To ensure reproducibility, I had to install these versions of the following packages specifically: Seurat_4.4.0, SeuratObject_4.1.4, SeuratData_0.2.2 (very specifically this commit “”, Azimuth_0.4.6.

As per this issue, https://github.com/satijalab/seurat-object/issues/165, if there is an error running sctransform, remove all relevant packages (and purge if using renv) and then install using the following method exactly, to build from source.

if (!requireNamespace(“remotes”, quietly = TRUE)) { install.packages(“remotes”) } options(repos = c(“https://satijalab.r-universe.dev”, getOption(“repos”))) remotes::install_version(“SeuratObject”, “4.1.4”) remotes::install_version(“Seurat”, “4.4.0”, upgrade = FALSE) # remotes will try to upgrade SeuratObject to v5, we need to tell it no

Licenses

The code in this analysis is covered by the MIT license and the written content on this website is covered by a Creative Commons CC-BY license.

Citations

Version Information

R version: R version 4.3.2 (2023-10-31)

Bioconductor version: 3.18


sessionInfo()
R version 4.3.2 (2023-10-31)
Platform: aarch64-apple-darwin20 (64-bit)
Running under: macOS Sonoma 14.5

Matrix products: default
BLAS:   /Library/Frameworks/R.framework/Versions/4.3-arm64/Resources/lib/libRblas.0.dylib 
LAPACK: /Library/Frameworks/R.framework/Versions/4.3-arm64/Resources/lib/libRlapack.dylib;  LAPACK version 3.11.0

locale:
[1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8

time zone: Australia/Melbourne
tzcode source: internal

attached base packages:
[1] stats     graphics  grDevices datasets  utils     methods   base     

other attached packages:
[1] workflowr_1.7.1

loaded via a namespace (and not attached):
 [1] jsonlite_1.8.8      compiler_4.3.2      BiocManager_1.30.22
 [4] renv_1.0.3          promises_1.2.1      Rcpp_1.0.12        
 [7] stringr_1.5.1       git2r_0.33.0        callr_3.7.3        
[10] later_1.3.2         jquerylib_0.1.4     yaml_2.3.8         
[13] fastmap_1.1.1       R6_2.5.1            knitr_1.45         
[16] tibble_3.2.1        rprojroot_2.0.4     bslib_0.6.1        
[19] pillar_1.9.0        rlang_1.1.3         utf8_1.2.4         
[22] cachem_1.0.8        stringi_1.8.3       httpuv_1.6.14      
[25] xfun_0.42           getPass_0.2-4       fs_1.6.3           
[28] sass_0.4.8          cli_3.6.2           magrittr_2.0.3     
[31] ps_1.7.6            digest_0.6.34       processx_3.8.3     
[34] rstudioapi_0.15.0   lifecycle_1.0.4     vctrs_0.6.5        
[37] evaluate_0.23       glue_1.7.0          whisker_0.4.1      
[40] fansi_1.0.6         rmarkdown_2.25      httr_1.4.7         
[43] tools_4.3.2         pkgconfig_2.0.3     htmltools_0.5.7