HNRNPH1 regulates the neuroprotective cold‐shock protein RBM3 expression through poison exon exclusion

A

Somal GFP intensity per unit area in GFP‐RBM3 i‐neurons transduced with lentivirus containing specific or non‐targeting sgRNA at 37 or 32°C (72 h) imaged by wide‐field microscopy. Only BFP‐positive (transduced) cells are included. Statistical analysis is performed between the specific and non‐targeting sgRNA within the temperature groups. Each data point represents one cell.

B

Median GFP intensity per unit area of GFP‐RBM3 i‐neurons at 37 or 32°C (72 h) treated with cycloheximide (CHX) at 50 μM for 72 h or actinomycin D (Act D) at 1 μM for 72 h.

C

Volcano plot showing differential expression analysis of all transcripts identified in i‐neurons at 37 and 32°C (72 h) from RNA‐Seq data.

D, E

qRT‐PCR of RBM3 Exon 3, Exon 4–5 (D) and pre‐mRNA (E) normalised to 18 s rRNA in i‐neurons at 37 and 32°C (72 h).

F, G

qRT‐PCR of RBM3 Exon 3, Exon 4–5 (F) and pre‐mRNA (G) normalised to 18 s rRNA in HeLa cells at 37 and 32°C (48 h).

H

Normalised RBM3 mRNA abundance (TPM) of control and selective regulator candidates knocked‐down K562 or HepG2 cells. Data are extracted from ENCODE project. 2 isogenic replicates are included in each condition.

I

qRT‐PCR of RBM3 mRNA normalised to GAPDH in control and HNRNPH1 KD HeLa cells at 37 or 32°C (48 h).

Figure EV2. Cooling and regulators involved in mRNA splicing change RBM3 transcript levels. Related to Fig 2 .