Hyperglycemia-induced overexpression of CREB3 L3 promotes the epithelial-to-mesenchymal transition in bladder urothelial cells in diabetes mellitus

Table 1 Antibody information used for western blot analysis

Antibody name	Manufacturers	CAS	Molecular weight (kDa)	Species	Dilution ratio
ACTIN	Servicebio	GB15003	42	Rabbit	1:1000
Occludin	Servicebio	GB111401	59	Rabbit	1:500
Claudin 1	servicebio	GB11032	20	Rabbit	1:500
CRP	ABCLONAL	A0224	25	Rabbit	1:500
CREB3 L3	Biorbyt	orb382910	49	Rabbit	1:500
N-cadherin	servicebio	GB111273	140	Rabbit	1:500
Vimentin	Servicebio	GB111308	54	Rabbit	1:500
E-cadherin	Servicebio	GB11082	135	Rabbit	1:500

CREB L3: CAMP-responsive element-binding protein 3 Like 3; CRP: C-reactive protein.

Table 2 Information on primers

Genes	Direction	Sequence (5’→3’)
CREB3 L3	Forward	ATCCTGGCAACTCTTGCTCC
CREB3 L3	Reverse	AGGTGATGCTGTTGCAGGTC
GAPDH	Forward	GAGTCAACGGATTTGGTCGT
GAPDH	Reverse	GACAAGCTTCCCGTTCTCAG

CREB L3: CAMP-responsive element-binding protein 3 Like 3; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

Table 3 Sequence of small interfering RNAs

Gene name	Sense sequence (5'--3')	Antisense sequence (5'--3')
si-NC	ACGUGACACGUUCGGAGAA	UUCUCCGAACGUGUCACGU
si-CREB3 L3-1	GGAAGAAGAAGAAGGAAUAUA	UAUUCCUUCUUCUUCUUCCUG
si-CREB3 L3-2	CGAGAAGAGUCUCCAGGAAGC	UUCCUGGAGACUCUUCUCGGG
si-CREB3 L3-3	GUACGAGUGUUCUCCAGAACU	UUCUGGAGAACACUCGUACAG

CREB L3: CAMP-responsive element-binding protein 3 Like 3; si-NC: Small interfering RNA negative control.

Table 4 Top 10 differentially upregulated and downregulated mRNAs involved in the inflammation pathways of bladder urothelium that are differentially expressed between patients with diabetic cystopathy and healthy controls by RNA sequencing analyses

Ensemble Gene ID	Gene name	Log2Fold change	P value	Regulation
ENSG00000173432	SAA1	9.14073	0.00000	Up
ENSG00000007171	NOS2	7.36349	0.00141	Up
ENSG00000102794	ACOD1	7.16781	0.00226	Up
ENSG00000163221	S100A12	7.06422	0.00001	Up
ENSG00000123838	C4BPA	6.64444	0.00071	Up
ENSG00000112115	IL17A	6.23562	0.01454	Up
ENSG00000164485	IL22RA2	6.02855	0.00162	Up
ENSG00000060566	CREB3 L3	5.75447	0.01491	Up
ENSG00000177455	CD19	5.62606	0.00004	Up
ENSG00000131203	IDO1	5.55962	0.00000	Up
ENSG00000006128	TAC1	-3.38685	0.00907	Down
ENSG00000143502	SUSD4	-2.17976	0.00717	Down
ENSG00000175344	CHRNA7	-2.17060	0.00039	Down
ENSG00000118432	CNR1	-2.08059	0.00014	Down
ENSG00000172458	IL17D	-2.01522	0.00173	Down
ENSG00000091831	ESR1	-1.97610	0.01013	Down
ENSG00000117335	CD46	-1.69696	0.00128	Down
ENSG00000132170	PPARG	-1.65880	0.01276	Down
ENSG00000107485	GATA3	-1.53300	0.00175	Down
ENSG00000168329	CX3CR1	-1.47299	0.02335	Down