Example data for multiWGCNA is stored in ExperimentHub. Access it like this:

# Load expression matrix and metadata
library(ExperimentHub)
## Loading required package: BiocGenerics
## Loading required package: generics
## 
## Attaching package: 'generics'
## The following objects are masked from 'package:base':
## 
##     as.difftime, as.factor, as.ordered, intersect, is.element, setdiff,
##     setequal, union
## 
## Attaching package: 'BiocGenerics'
## The following objects are masked from 'package:stats':
## 
##     IQR, mad, sd, var, xtabs
## The following objects are masked from 'package:base':
## 
##     Filter, Find, Map, Position, Reduce, anyDuplicated, aperm, append,
##     as.data.frame, basename, cbind, colnames, dirname, do.call,
##     duplicated, eval, evalq, get, grep, grepl, is.unsorted, lapply,
##     mapply, match, mget, order, paste, pmax, pmax.int, pmin, pmin.int,
##     rank, rbind, rownames, sapply, saveRDS, table, tapply, unique,
##     unsplit, which.max, which.min
## Loading required package: AnnotationHub
## Loading required package: BiocFileCache
## Loading required package: dbplyr
## Registered S3 method overwritten by 'bit64':
##   method          from 
##   print.bitstring tools
eh = ExperimentHub()
eh_query = query(eh, c("multiWGCNAdata"))

## download the autism data and metadata
autism_se = eh_query[["EH8219"]]
## see ?multiWGCNAdata and browseVignettes('multiWGCNAdata') for documentation
## loading from cache
## require("SummarizedExperiment")

Now, proceed with the multiWGCNA analysis:

# Load multiWGCNA R package
library(multiWGCNA)
## Loading required package: ggalluvial
## Loading required package: ggplot2
# Obtain metadata
sampleTable = colData(autism_se)

# Randomly sample 2000 genes from the expression matrix
set.seed(1)
autism_se = autism_se[sample(rownames(autism_se), 2000),]

# Check the data
assays(autism_se)[[1]][1:5, 1:5]
##              GSM706412 GSM706413 GSM706414 GSM706415 GSM706416
## ILMN_1672121 11.034264 10.446682 11.473705 11.732849  11.43105
## ILMN_2151368 10.379812  9.969130  9.990030  9.542288  10.26247
## ILMN_1757569  9.426955  9.050024  9.347505  9.235251   9.38837
## ILMN_2400219 12.604047 12.886037 12.890658 12.446960  12.98925
## ILMN_2222101 12.385019 12.748229 12.418027 11.690253  13.10915
sampleTable
## DataFrame with 58 rows and 3 columns
##                Sample      Status      Tissue
##           <character> <character> <character>
## GSM706412   GSM706412      autism          FC
## GSM706413   GSM706413      autism          FC
## GSM706414   GSM706414      autism          FC
## GSM706415   GSM706415      autism          FC
## GSM706416   GSM706416      autism          FC
## ...               ...         ...         ...
## GSM706465   GSM706465    controls          TC
## GSM706466   GSM706466    controls          TC
## GSM706467   GSM706467    controls          TC
## GSM706468   GSM706468    controls          TC
## GSM706469   GSM706469    controls          TC
# Set the alpha level for statistical analyses and the soft power for network construction
alphaLevel = 0.05
softPower = 10

# If your sample traits include numbers that you'd like to be considered numerical 
# variables rather than categorical variables, set detectNumbers = TRUE
detectNumbers = FALSE

We now perform network construction, module eigengene calculation, module-trait correlation.

# Define our conditions for trait 1 (disease) and 2 (brain region)
conditions1 = unique(sampleTable[,2])
conditions2 = unique(sampleTable[,3])
# Construct the combined networks and all the sub-networks (autism only, controls only, FC only, and TC only)
# Same parameters as Tommasini and Fogel. BMC Bioinformatics
myNetworks = constructNetworks(autism_se, sampleTable, conditions1, conditions2, 
                                  networkType = "signed", TOMType = "unsigned", 
                                  power = softPower, minModuleSize = 100, maxBlockSize = 25000,
                                  reassignThreshold = 0, minKMEtoStay = 0, mergeCutHeight = 0,
                                  numericLabels = TRUE, pamRespectsDendro = FALSE, 
                                  deepSplit = 4, verbose = 3)

Carry on with the multiWGCNA analysis according to the generalWorkflow.Rmd vignette!

sessionInfo()
## R version 4.6.0 RC (2026-04-17 r89917)
## Platform: x86_64-pc-linux-gnu
## Running under: Ubuntu 24.04.4 LTS
## 
## Matrix products: default
## BLAS:   /home/biocbuild/bbs-3.24-bioc/R/lib/libRblas.so 
## LAPACK: /usr/lib/x86_64-linux-gnu/lapack/liblapack.so.3.12.0  LAPACK version 3.12.0
## 
## locale:
##  [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C              
##  [3] LC_TIME=en_GB              LC_COLLATE=C              
##  [5] LC_MONETARY=en_US.UTF-8    LC_MESSAGES=en_US.UTF-8   
##  [7] LC_PAPER=en_US.UTF-8       LC_NAME=C                 
##  [9] LC_ADDRESS=C               LC_TELEPHONE=C            
## [11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C       
## 
## time zone: America/New_York
## tzcode source: system (glibc)
## 
## attached base packages:
## [1] stats4    stats     graphics  grDevices utils     datasets  methods  
## [8] base     
## 
## other attached packages:
##  [1] multiWGCNA_1.9.3            ggalluvial_0.12.6          
##  [3] ggplot2_4.0.2               SummarizedExperiment_1.41.1
##  [5] Biobase_2.71.0              GenomicRanges_1.63.2       
##  [7] Seqinfo_1.1.0               IRanges_2.45.0             
##  [9] S4Vectors_0.49.2            MatrixGenerics_1.23.0      
## [11] matrixStats_1.5.0           multiWGCNAdata_1.9.0       
## [13] ExperimentHub_3.1.0         AnnotationHub_4.1.0        
## [15] BiocFileCache_3.1.0         dbplyr_2.5.2               
## [17] BiocGenerics_0.57.1         generics_0.1.4             
## [19] BiocStyle_2.39.0           
## 
## loaded via a namespace (and not attached):
##   [1] RColorBrewer_1.1-3    rstudioapi_0.18.0     jsonlite_2.0.0       
##   [4] magrittr_2.0.5        farver_2.1.2          rmarkdown_2.31       
##   [7] vctrs_0.7.3           memoise_2.0.1         base64enc_0.1-6      
##  [10] htmltools_0.5.9       S4Arrays_1.11.1       dynamicTreeCut_1.63-1
##  [13] curl_7.0.0            SparseArray_1.11.13   Formula_1.2-5        
##  [16] sass_0.4.10           bslib_0.10.0          htmlwidgets_1.6.4    
##  [19] httr2_1.2.2           impute_1.85.0         cachem_1.1.0         
##  [22] igraph_2.2.3          lifecycle_1.0.5       iterators_1.0.14     
##  [25] pkgconfig_2.0.3       Matrix_1.7-5          R6_2.6.1             
##  [28] fastmap_1.2.0         digest_0.6.39         colorspace_2.1-2     
##  [31] patchwork_1.3.2       AnnotationDbi_1.73.1  Hmisc_5.2-5          
##  [34] RSQLite_2.4.6         filelock_1.0.3        httr_1.4.8           
##  [37] polyclip_1.10-7       abind_1.4-8           compiler_4.6.0       
##  [40] rngtools_1.5.2        bit64_4.6.0-1         withr_3.0.2          
##  [43] doParallel_1.0.17     htmlTable_2.4.3       S7_0.2.1-1           
##  [46] backports_1.5.1       viridis_0.6.5         DBI_1.3.0            
##  [49] ggforce_0.5.0         MASS_7.3-65           rappdirs_0.3.4       
##  [52] DelayedArray_0.37.1   flashClust_1.1-4      tools_4.6.0          
##  [55] foreign_0.8-91        otel_0.2.0            nnet_7.3-20          
##  [58] glue_1.8.1            grid_4.6.0            checkmate_2.3.4      
##  [61] cluster_2.1.8.2       gtable_0.3.6          tzdb_0.5.0           
##  [64] preprocessCore_1.73.0 tidyr_1.3.2           hms_1.1.4            
##  [67] data.table_1.18.2.1   WGCNA_1.74            tidygraph_1.3.1      
##  [70] XVector_0.51.0        ggrepel_0.9.8         BiocVersion_3.23.1   
##  [73] foreach_1.5.2         pillar_1.11.1         stringr_1.6.0        
##  [76] splines_4.6.0         dplyr_1.2.1           tweenr_2.0.3         
##  [79] lattice_0.22-9        survival_3.8-6        bit_4.6.0            
##  [82] tidyselect_1.2.1      Biostrings_2.79.5     knitr_1.51           
##  [85] gridExtra_2.3         bookdown_0.46         xfun_0.57            
##  [88] graphlayouts_1.2.3    stringi_1.8.7         yaml_2.3.12          
##  [91] evaluate_1.0.5        codetools_0.2-20      ggraph_2.2.2         
##  [94] tibble_3.3.1          BiocManager_1.30.27   cli_3.6.6            
##  [97] rpart_4.1.27          jquerylib_0.1.4       dichromat_2.0-0.1    
## [100] Rcpp_1.1.1-1          png_0.1-9             fastcluster_1.3.0    
## [103] parallel_4.6.0        readr_2.2.0           blob_1.3.0           
## [106] dcanr_1.27.0          doRNG_1.8.6.3         viridisLite_0.4.3    
## [109] scales_1.4.0          purrr_1.2.2           crayon_1.5.3         
## [112] rlang_1.2.0           cowplot_1.2.0         KEGGREST_1.51.1