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library(scRNAseq)
library(SingleCellExperiment)
library(scran)
library(scater)
library(Seurat)
Warning in fun(libname, pkgname): rgeos: versions of GEOS runtime 3.10.3-CAPI-1.16.1
and GEOS at installation 3.9.1-CAPI-1.14.2differ
library(tibble)
library(dplyr)

source(here::here("code", "data-analysis-utils.R"))
zeisel <- ZeiselBrainData()
snapshotDate(): 2021-10-19
see ?scRNAseq and browseVignettes('scRNAseq') for documentation
loading from cache
see ?scRNAseq and browseVignettes('scRNAseq') for documentation
loading from cache
see ?scRNAseq and browseVignettes('scRNAseq') for documentation
loading from cache
snapshotDate(): 2021-10-19
see ?scRNAseq and browseVignettes('scRNAseq') for documentation
loading from cache
colLabels(zeisel) <- zeisel$level1class
table(zeisel$tissue)

ca1hippocampus       sscortex 
          1314           1691 
table(zeisel$level1class)

astrocytes_ependymal    endothelial-mural         interneurons 
                 224                  235                  290 
           microglia     oligodendrocytes        pyramidal CA1 
                  98                  820                  939 
        pyramidal SS 
                 399 
zeisel <- logNormCounts(zeisel)
zeisel <- runPCA(zeisel)
zeisel <- runTSNE(zeisel, dimred = "PCA")
plotTSNE(zeisel, colour_by = "level1class")

plotTSNE(zeisel, colour_by = "level2class")

mgs <- findMarkers(zeisel, test.type = "wilcox", pval.type = "all")
oligo_mgs <- mgs$oligodendrocytes %>% 
  as.data.frame() %>% 
  rownames_to_column("gene") %>% 
  rowid_to_column() %>% 
  as_tibble() %>% 
  print(n = 20)
# A tibble: 20,006 × 11
   rowid gene     p.value      FDR summary.AUC AUC.astrocytes_… AUC.endothelial…
   <int> <chr>      <dbl>    <dbl>       <dbl>            <dbl>            <dbl>
 1     1 Plp1    3.79e-58 3.17e-54       0.996            0.989            0.995
 2     2 Sept4   4.59e-58 3.17e-54       0.996            0.979            0.957
 3     3 Mbp     4.75e-58 3.17e-54       0.996            0.988            0.998
 4     4 Cnp     7.28e-58 3.64e-54       0.995            0.984            0.997
 5     5 Mobp    1.35e-57 5.39e-54       0.994            0.987            0.993
 6     6 Cryab   3.76e-57 1.25e-53       0.992            0.965            0.984
 7     7 Mog     4.37e-57 1.25e-53       0.991            0.988            0.993
 8     8 Dbndd2  6.00e-57 1.50e-53       0.991            0.965            0.988
 9     9 Ugt8a   8.91e-57 1.98e-53       0.990            0.983            0.993
10    10 Gsn     7.47e-55 1.50e-51       0.981            0.972            0.972
11    11 Taldo1  1.44e-54 2.62e-51       0.980            0.976            0.985
12    12 Tmem88b 4.68e-54 7.80e-51       0.978            0.956            0.968
13    13 Ermn    7.47e-54 1.15e-50       0.977            0.971            0.976
14    14 281046… 9.43e-54 1.35e-50       0.976            0.968            0.981
15    15 Tspan2  2.21e-52 2.95e-49       0.970            0.972            0.983
16    16 Cldn11  2.94e-52 3.67e-49       0.970            0.971            0.984
17    17 Hmgcs1  9.40e-52 1.11e-48       0.830            0.830            0.970
18    18 Qdpr    1.02e-51 1.14e-48       0.967            0.964            0.975
19    19 Sept7   1.91e-51 2.01e-48       0.966            0.909            0.902
20    20 Mag     2.15e-51 2.16e-48       0.965            0.958            0.978
# … with 19,986 more rows, and 4 more variables: AUC.interneurons <dbl>,
#   AUC.microglia <dbl>, AUC.pyramidal.CA1 <dbl>, AUC.pyramidal.SS <dbl>
oligo_mgs
# A tibble: 20,006 × 11
   rowid gene    p.value      FDR summary.AUC AUC.astrocytes_e… AUC.endothelial…
   <int> <chr>     <dbl>    <dbl>       <dbl>             <dbl>            <dbl>
 1     1 Plp1   3.79e-58 3.17e-54       0.996             0.989            0.995
 2     2 Sept4  4.59e-58 3.17e-54       0.996             0.979            0.957
 3     3 Mbp    4.75e-58 3.17e-54       0.996             0.988            0.998
 4     4 Cnp    7.28e-58 3.64e-54       0.995             0.984            0.997
 5     5 Mobp   1.35e-57 5.39e-54       0.994             0.987            0.993
 6     6 Cryab  3.76e-57 1.25e-53       0.992             0.965            0.984
 7     7 Mog    4.37e-57 1.25e-53       0.991             0.988            0.993
 8     8 Dbndd2 6.00e-57 1.50e-53       0.991             0.965            0.988
 9     9 Ugt8a  8.91e-57 1.98e-53       0.990             0.983            0.993
10    10 Gsn    7.47e-55 1.50e-51       0.981             0.972            0.972
# … with 19,996 more rows, and 4 more variables: AUC.interneurons <dbl>,
#   AUC.microglia <dbl>, AUC.pyramidal.CA1 <dbl>, AUC.pyramidal.SS <dbl>
oligo_mgs %>% 
  filter(gene == "Olig2")
# A tibble: 1 × 11
  rowid gene   p.value      FDR summary.AUC AUC.astrocytes_epe… AUC.endothelial…
  <int> <chr>    <dbl>    <dbl>       <dbl>               <dbl>            <dbl>
1   315 Olig2 7.19e-12 4.57e-10       0.694               0.667            0.726
# … with 4 more variables: AUC.interneurons <dbl>, AUC.microglia <dbl>,
#   AUC.pyramidal.CA1 <dbl>, AUC.pyramidal.SS <dbl>
plotExpression(zeisel, "Olig2", x = "level1class")

oligo_mgs %>% 
  filter(gene == "Ptgds")
# A tibble: 1 × 11
  rowid gene   p.value      FDR summary.AUC AUC.astrocytes_epe… AUC.endothelial…
  <int> <chr>    <dbl>    <dbl>       <dbl>               <dbl>            <dbl>
1   146 Ptgds 3.40e-22 4.65e-20       0.796               0.844            0.854
# … with 4 more variables: AUC.interneurons <dbl>, AUC.microglia <dbl>,
#   AUC.pyramidal.CA1 <dbl>, AUC.pyramidal.SS <dbl>
zeisel <- readRDS(here::here("data", "real_data", "zeisel.rds"))
plotTSNE(zeisel, colour_by = "label") 

plotUMAP(zeisel, colour_by = "label") 

plot_cluster_counts(zeisel)

plot_cluster_mean_boxplot(zeisel)


devtools::session_info()
─ Session info  ──────────────────────────────────────────────────────────────
 hash: weary face, Mrs. Claus: light skin tone, man student: medium-light skin tone

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 language (EN)
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 pandoc   2.18 @ /apps/easybuild-2022/easybuild/software/MPI/GCC/11.3.0/OpenMPI/4.1.4/RStudio-Server/2022.07.2+576-Java-11-R-4.1.2/bin/pandoc/ (via rmarkdown)

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 [1] /home/jpullin/R/x86_64-pc-linux-gnu-library/4.1
 [2] /apps/easybuild-2022/easybuild/software/MPI/GCC/11.3.0/OpenMPI/4.1.4/R/4.1.2/lib64/R/library

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