TABLE 1. Monolayer and anchorage independent growth of NSCLC cell lines with knockdown of RAL GTPase.

Cells were transfected with siRNA against RAL GTPase and plated for proliferation and anchorage independent growth assay (see Materials and Methods). Monolayer growth was measured 4 days after transfection with results representing the mean of 2-3 studies with 4-5 replicates per cell line. Anchorage independent growth results are mean of three experiments.

			RALA Knockdown		RALB Knockdown		RALA+B Knockdown
			Monolayer Growth	Anchorage Independent Growth	Monolayer Growth	Anchorage Independent Growth	Monolayer Growth	Anchorage Independent Growth
Cell Line	Histology	KRAS
H322	Adenocarcinoma	WT	−52%*	−41.2%*	−33%*	−42.3%*	−46%*	−39%*
H1703	Adenocarcinoma	WT	N.S.	N.S.	N.S.	N.S.	N.S.	−52.3%*
H2228	Adenocarcinoma	WT	N.S.	N.S.	N.S.	N.S.	N.S.	N.S.
H292	Mucoepidermoid	WT	N.S.	No Colony	−23%*	No Colony	N.S.	No Colony
H4006	Adenocarcinoma	WT	N.S.	No Colony	N.S.	No Colony	N.S.	No Colony
Calu3	Adenocarcinoma	WT	−33%*	No Colony	−36%*	No Colony	−39%*	No Colony
H2122	Adenocarcinoma	G12C	−91%*	−97%*	−61%*	−89.2%*	−42%*	−79.6%*
H358	Adenocarcinoma	G12C	−24%*	−38%*	N.S.	−27.6%*	−86%*	−83.4%*
A549	Adenocarcinoma	G12S	−26%*	−24.5%*	−29%*	N.S.	−27%*	−51.3%*
H2009	Adenocarcinoma	G12A	−25%*	−35%*	−19%*	−28.7%*	28%*	−15.8%*
H157	Squamous Cell	G12R	−24%*	N.S.	−19%*	N.S.	N.S.	N.S.
SW1573	Alveolar Cell	G12A	N.S.	No Colony	N.S.	No Colony	N.S.	No Colony

^*P<0.05 (student's t-test)

(+) indicates increase compared to control

(−) indicates decrease compared to control

“N.S.” is no significant change

“No Colony” indicates cells did not exhibit anchorage independent growth.